Procedure_Catalogue-Phoenix-All_Cobined.pdf - Bio Group

PRODUCT PACK SIZE PAGE NO.

ACID PHOSPHATASE 1ALKALINE PHOSPHATASE 2CHOLESTEROL 3CHOLINESTERASE 4CK‐MB 5 x 2.5mL 5CK‐NAC 5 x 2.5mL 6GLUCOSE 7SGOT 8SGPT 9TRIGLYCERIDES 10UREA‐B 11UREA U.V 12URIC ACID 13

ALKALINE PHOSPHATASE (S.L) 14ALPHA AMYLASE (S.L) 15CHOLESTEROL(S.L) 16CHOLINESTERASE ( S.L) 17CK‐MB (S.L) 18CK‐NAC (S.L) 19ENZYMATIC CREATININE (S.L) 20GAMMA GT (S.L) 21GLUCOSE (S.L) 22HDL ‐ C DIRECT 23LDH ‐ P (S.L) 24LDL ‐ C DIRECT 25LIPASE (S.L) 26SGOT (S.L) 27SGPT (S.L) 28TRIGLYCERIDES (S.L) 29UREA U.V (S.L) 30URIC ACID (S.L) 31

ALBUMIN 32BILIRUBIN DIRECT 33BILIRUBIN TOTAL ‐TAB 34BILIRUBIN TOTAL 35BILIRUBIN TOTAL& DIRECT 36CALCIUM 37CALCIUM (ARSENAZO) 38CHLORIDE 39COPPER 2 x 25mL 40CREATININE 41HAEMOGLOBIN 42HDL‐CHOLESTEROL 43INORGANIC PHOSPHOROUS 44IRON 45MAGNESIUM 46MICROPROTEIN 47TIBC 50T 48TOTAL PROTEIN 49ZINC 2 x 10mL 50

ALPHA1ACID GLYCOPROTEIN WITH CALIBRATOR 51APOA1 WITH CALIBRATOR 1 x 30ml/1 x 5ml/1ml 52APOB WITH CALIBRATOR 1 x 30ml/1 x 5ml/1ml 53

Lypho CHEK5 x 2 mL

10 x 3 mL 4 x 50 mL /4 x 100 mL

5 x 3 mL

4 x 100 mL /4 x 250 mL/ 4 x 500 mL 5 x 20 mL/4 x 50 mL 5 x 20 mL/4 x 50 mL 5 x 20mL / 4 x 50 mL

200 mL 4 x 50 mL / 4 x 100 mL 5 x 20 mL/ 4 x 50 mL

Liqui CHEK2 x 10 mL /2 x 30 mL/2 x 50 mL

4 x 5 mL /4 x 10 mL5 x 25 mL / 5 x 100 mL/ 4 x 250 mL

2 x 10 mL2 x 12.5 mL/2 x 20 mL2 x 10 mL /2 x 30 mL

2 x 40 mL /2 x 60 mL/4 x 60 mL2 x 10 mL / 2 x 30 mL

5 x 100 mL /1 x 1000 mL2 x 40 mL /2 x 60 mL / 4 x 60 mL

2 x 10 mL /2 x 30 mL2 x 40 mL /2 x 60 mL

1 x 25 mL3 x 20 mL /3 x 50 mL / 4 x 125 mL 3 x 20 mL / 3 x 50 mL/ 4 x 125 mL

4 x 10 mL /5 x 25 mL / 6 x 50 mL/ 5 x 100 mL2 x 30 mL / 2 x 50 mL / 2 x 125 mL2 x 30 mL / 2 x 50 mL / 2 x 100 mL

ChemCHEK4 x 50 mL4 x 50 mL4 x 50 mL4 x 50 mL4 x 50 mL

4 x 25 mL / 4 x 50 mL4 x 25 mL4 x 50 mL

4 x 50 mL1000 mL

4 x 25 mL4 x 25 mL / 4 x 50 mL

2 x 50 mL4 x 25 mL1 x 50 mL

4 x 50 mL

SensIT1 x 20 mL/1 x 10ml/1ml

PRODUCT PACK SIZE PAGE NO.ASO LATEX WITH CALIBRATOR 1 x 45ml/1 x 5 ml/1ml 54ASO LEIT WITH CALIBRATOR 1 x 24ml/1 x 8 ml/1ml, 2 x 24ml/2 x 8 ml/2ml 55C3 WITH CALIBRATOR 1 x 30ml/1 x 5ml/1ml 56C4 WITH CALIBRATOR 1 x 30ml/1 x 5ml/1ml 57CERULOPLASMIN WITH CALIBRATOR 1 x 30ml/1 x 5ml/1ml 58CRP LATEX WITH CALIBRATOR 1 x 45ml/1 x 5 ml/2ml 59CRP LEIT WITH CALIBRATOR 1 x 24ml/1 x 8 ml/2ml, 2 x 24ml/2 x 8 ml/2ml 60CRP ULTRA WITH CALIBRATOR 1 x 18ml/1 x 9ml/2ml 61CYSTATIN C WITH CALIBRATOR 1 x 25ml/1 x 5ml/2ml 62FERRITIN+CALIBRATOR 1 x 30ml/1 x 10ml/1ml 63HBA1C DIRECT WITH CALIBRATOR 64

1 x 30ml/1 x 10ml/1ml 65 1 x 15ml/1 x 15ml/1ml 66 1 x 30ml/1 x 10ml/1ml 67 1 x 32ml/1 x 8ml/1ml 68

LIPOPROTEIN (A) WITH CALIBRATOR 1 x 20mL/1 x 4mL/1mL 69MICRO ALBUMIN LATEX WITH CALIBRATOR 70MICRO ALBUMIN IT WITH CALIBRATOR 2 x 25mL/2 x 5mL/1mL 71PREALBUMIN WITH CALIBRATOR 72RF LATEX WITH CALIBRATOR 1 x 45ml/1 x 5 ml/2ml 73R.F LEIT WITH CALIBRATOR 1 x 24ml/1 x 8 ml/1ml, 2 x 24ml/2 x 8 ml/2ml 74TRANSFFERIN WITH CALIBRATOR 75

ASO 50 T / 100 T 76CRP 50 T / 100 T 77RF 50 T / 100 T 78RPR 50 T /125 T / 500 T 79BLOOD GROUPINGANTI‐A 80

3 x 10mL 813 x 10mL 82

ANTI‐B 8384

1 x 10 ml 85

PT (SL) 86APTT 87

DILUENT 10 Litre /20 Litre 88E‐Z CLEANER 89LYSE 90PROBE CLEANER 91RINSE 10 Litre /20 Litre 92CONTROLS & CALIBRATORSASO CALIBRATOR 93MICROALBUMIN CALIBRATOR 94CRP CALIBRATOR 95

96PROTEIN CALIBRATOR 1 x 1 97MULTICALIBRATOR 98FERRITIN CALIBRATOR 99HbA1c DIRECT MULTI CALIBRATOR 100RF CALIBRATOR 101HbA1c CONTROL LEVEL 1 &2 2 x 0.5ml 102PROTEIN CONTROL(IMMUNOLOGY) 103QUALICHECK NORM & PATH 104

4 x 7.5/1 x 9.5/1 x .5/4 x .5 mLIgM WITH CALIBRATORIgG WITH CALIBRATORIgA WITH CALIBRATORIgE WITH CALIBRATOR

1 x 45 mL/1 x 5mL/1mL

1 x 25 mL/1 x 5mL/1mL

1 x 30 mL/1 x 10mL/1mLSero CHEK

1 x 10 mLANTI‐A,B&D(IgG+IgM)ANTI‐A,B&D(IgM)

1 x 10 mLANTI‐D (IgG+IgM) 1 x 10 mLANTI‐D (IgM)CoagTHREE

2 x 4 mL2 x 4 mL

HaemoCHEK

4 x 50 mL500 mL /2 x 500 mL

4 x 50 mL

1 x 1 mL1 x 1 mL1 x 2 mL

Lp(a) CALIBRATOR 1 x 1 mL

5 x 3 mL1 x 1 mL

4 x 0.5 mL1 x 1 mL

1 x 1 mL2 x 5 mL

PRODUCT PACK SIZE PAGE NO.

ALPHA 1‐ACID GLYCOPROTEIN 105C3 1 x 35mL /1 x6mL 107C4 109APO A1 1 x 35/1x6mL 111APO B 113CERULOPLASMIN 115PREALBUMIN 117TRANSFERRIN 1 x 25mL /1 x 4mL 119

121123

1 x 30mL/1 x 10mL 1251 x 22mL/1 x 6.5mL 127

FERRITIN 1 x 20mL/1 x 11mL 129CYSTATIN‐C 131HbA1c DIRECT 1 x 30/1 x 9.5/1 x 0.5/2x63mL 133ASO 135CRP 137RF 139

141MICROALBUMIN 143CRP Ultra 145ALBUMIN 148ALKALINE PHOSPHATASE 150AMYLASE 152BILIRUBIN DIRECT 154BILIRUBIN TOTAL (TAB) 156CALCIUM (ARSENAZO) 158CHLORIDE 160CHOLESTEROL 162CREATININE 164CREATINE KINASE 166ENZYMATIC CREATININE 168GAMMA GT 170GLUCOSE 172HDL – C DIRECT 174LDH 176LDL – C DIRECT 178MAGNESIUM 180PHOSPHOROUS 182SGOT 184SGPT 186TOTAL PROTEIN 188TRIGLYCERIDES 190UREA U V 192URIC ACID 194

Mispa Nano1 x 25mL /1 x4 mL

1 x35 mL/1x6mL

1 x 35mL/1 x 6 mL1 x 35 mL/1 x6 mL1 x25mL /1 x 4 mL

IgM 1 x 30/1 x 10 mLIgG 1 x 15mL/1 x 15 mLIgAIgE

1 x 23mL/1 x 5.5 mL

1 x 20/1 x 11 mL1 x 20/1 x8 mL1x 20/1 x 8 mL

Lp (a) 1 x 23/1 x 5.5 mL1 x 23/1 x 5.5 mL1x20 / 1x11 mL

4 x 30 mL2 x 30 / 2 x 8 mL

2 x 30 mL4 x 30 / 2 x 8 mL4 x 35 / 2 x 10 mL

2 x 35 mL2 x 30 mL4 x 35 mL

4 x 35 / 2 x 18 mL1 x 30 / 1 x 8 mL2 x 35 / 2 x 12 mL1 x 30 / 1 x 8 mL

4 x 35 mL2 x 30 / 2 x 10 mL1 x 30 / 1 x 8 mL1 x 30 / 1 x 10 mL

1 x 30 mL1 x 30 mL

4 x 35 / 2 x 18 mL4 x 35 / 2 x 18 mL

4 x 30 mL4 x 35 mL

4 x 30 / 2 x 16 mL2 x 35 mL

INTENDED USE

This reagent is intended for in vitro quantitative determination of Acid phosphatasein serum.

- Alpha - naphthylphosphate method

- Linear upto 150 U/L

- Tartrate included for determination of non-prostatic acid phosphatase

CLINICAL SIGNIFICANCE

Acid phosphatase is present in lysosomes, which are organells present in all cells withthe possible exception of erythrocytes. The greatest concentrations of ACP activityoccurs in liver, spleen, erythrocytes, platelets, bone marrow & the prostate gland whichis the richest source and it contributes a small portion of the enzyme present in serafrom healthy males.

Determination of ACP activity in serum is almost directed toward the prostatic enzymewith the intent of detecting or monitoring carcinoma of the prostate. Elevation of theenzymatic activity of prostatic ACP & thus of total ACP activity are found in the seraof about 60% of men with prostatic cancer & metastase.

Slight or moderate elevations in total ACP activity often occur in Pagets disease inhyperparathyroidism with skeletal involvement & in the presence of malignant invasionof the bones by cancers, such as breast cancer in women, unlike prostatic ACP, in thesecases the serum ACP activity is not inhibited by tartrate.

PRINCIPLE

Acid Phosphatase activity present in the sample is determined according to thefollowing reactions.

Acid Phosphatase

alpha - naphthyl-phosphate+H2O -----------------------> alpha –naphthol + phosphate

a-naphthol+ Fast Red TR -------------> Azo dye

Tartrate is used as specific inhibitor of the prostatic fraction.

REAGENT COMPOSITION

ACID PHOSPHATASE R1 5 x 2 mL

Citrate Buffer (pH 5.2) 50 mmol/L

ACID PHOSPHATASE R2 (Tablets) 1 x 5 x 2 mL

alpha–naphtylphosphate 10 mmol / L

Fast red TR 6 mmol / L

ACID PHOSPHATASE R3 1 x 1 mL

Sodium tartrate 2 mmol/L

STORAGE AND STABILITY

The sealed reagents are stable upto the expiry date stated on the label, when storedat 2 - 80C.

LINEARITY

This reagent is linear upto 150 U/L

If the concentration is greater than linearity (150 U/L), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

REFERENCE RANGE

It is recommended that each laboratory establish its own reference values.

The following value may be used as guide line.

Total acid phosphatase:

Men : < 5.4 U/L

Women : < 4.2 U/L

Prostatic acid Phosphatase : < 1.7 U/L

PREPARATION AND STABILITY OF WORKING REAGENT

Dissolve one tablet (R2) with 2 mL of Reagent 1 (R1)

The working reagent is stable for 2 days at 2-80C.

Wait for 10 minutes for complete dissolution.

PRECAUTION

To avoid contamination, use clean laboratory wares.

Avoid direct exposure of working reagent to light.

Acid phosphatase is extremely temperature labile. The sample should therefore becentrifugated immediately after coagulation. The serum should be cooled and proceedas quickly as possible. Do not use hemolytic serum. If the serum is to be examinedafter some time, its pH value should be adjusted to about 5 (0.02 mL acetate buffer5 M per 1 mL of serum).

SAMPLE

5 x 2 mL

11201001ACID PHOSPHATASE

GENERAL SYSTEM PARAMETER

Mode of Reaction Kinetic

Slope of reaction Increasing

Wavelength 405 nm

Temperature 370C

Factor 750

Linearity 150 U/L

Blank DI Water

Delay 300 sec.

No of reading 3

Interval 60 sec

Sample volume 100 µL

Reagent volume 1000 µL

Cuvette 1 cm light path

LABORATORY PROCEDURE

Total non inhibitor by tartrate fraction

Working reagent 1000 µL 1000 µL

Tartrate Solution R3

- 10 µL

Sample 100 µL 100 µL

Mix, and incubate for 5 minutes at 370C. Measure the change in absorbance perminute ( OD/min) during 3 minutes.

CALCULATION

Total acid phosphatase (U/L) = 750 x ( OD/min.) of total

Non prostatic ACP activity (U/L) = 750 x ( OD/min.) of non inhibitor fraction.

Fraction of prostatic acid phosphatase(U/L) = Total ACP activity - Non-prostatic ACP activity.

BIBLIOGRAPHY

1 Hillman G. Z., Clin.Chem. Biochem 9.273 (1971).

ADL/V.02/February 2013

Fresh serum only. Do not use plasma.

1

INTENDED USE

This reagent is intended for in vitro quantitative determination of Alkaline Phosphatasein serum or plasma.

- DGKC – SCE recommended procedure with DEA buffer


- Working reagent stable upto 7 days at 2-80C


Alkaline phosphatase is widely distributed throughout the body, but clinicallyimportant one for diagnostic reasons are in bone, liver, placenta & intestine. Growingbone is associated with the release of ALP and so in childhood the level of ALP isaround 3 times of that of adult. During pregnancy in 2nd&3rd trimester the enzyme risesconsiderably due to placenta releasing ALP. It can be used to examine placentalfunction.

Elevated levels are seen in bone diseases, e.g. pagets disease, rickets, osteoblasticmetastatic & in obstructive disease of biliary tract.

Decreased levels are rarely seen. e.g. in Vitamin A resistant rickets.

PRINCIPLE

Kinetic determination of ALP according to the following reaction

ALP

p-nitrophenyl phosphate + H2O-------- >p-nitrophenol+Inorgnic phosphate

ALP = Alkaline Phosphatase

REAGENT COMPOSITION

ALKALINE PHOSPHATASE R1 1 x 33 mL

Diethanolamine Buffer 1 mmol/L

Magnesium Chloride 0.5 mmol/L


P-Nitrophenyl phosphate 10 mmol/L


The sealed reagents are stable upto the expiry date stated on the label, when storedat 2-80C.

LINEARITY

The reagent is linear, up to 900 U/L.

If the concentration is greater than linearity (900 U/L), dilute the sample with normalsaline & repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE

It is recommended that each laboratory establish its own reference values. Thefollowing value may be used as guide line.

Adults : 100 – 290 U/L

Children : 180- 1200 U/L


Reconstitute the Reagent 2(R2) with the volume Reagent 1(R1) mentioned on the viallabel. The reconstituted reagent is stable for 7 days at 2-80C.

NOTE

Discard the working reagent if the blank absorbance exceeds 1.0 at 405 nm.

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure of workingreagent to light.

SAMPLE

Serum / plasma (free of haemolysis)

ALKALINE PHOSPHATASE10 x 3 mL

11202001




Wavelength 405 nm

Temperature 370C

Factor 2750

Blank DI water

Linearity 900 U/L

Delay time 60 sec

No of readings 3

Interval 60 sec





Working reagent 1000 µL

Sample 2µL

Mix and incubate at 370C for one minute. Measure the change in absorbance perminute ( OD/min) during 3 minutes.

CALCULATION

ALP Activity (U/L) = ( OD/ min.) x 2750

BIBLIOGRAPHY

1. Scand J. Clin. Lab. Invest. 32:29 (1974)2. Tietz, N.W..; Clin. Chem. 29:751 (1983)3. Z Klin. Chem. U. Clin. Biochem. 8 (1970) 658; 10 (1972), 182.


2

INTENDED USE

This reagent is intended for in vitro quantitative determination of Cholesterol in serumor plasma.

- Cholesterol Oxidase Peroxidase methodology

- Linear up to 500 mg/dL

- Reconstituted reagent stable up to 90 days at 2- 80C

- Lipemic clearing system, minimizes reruns


It is the main lipid found in the blood, bile & brain tissues. It is also one of the mostimportant steroids of the body & is a precursor of many steroid hormones. Twothirds of cholesterol present in the blood is esterified. The liver metabolizes thecholesterol & it is transported in the blood stream by lipoproteins.

Increased levels are found in hypercholesterolemia, hyperlipidemia, hypothyroidism,uncontrolled diabetes, nephritic syndrome & cirrhosis.

Decreased levels are found in malabsorption, malnutrition, hyperthyroidism,anaemia & liver diseases.

PRINCIPLE

Enzymatic colorimetric determination of total cholesterol according to the followingreactions.

Cholesterol esterase

Cholesterol ester +H2O ---------------------------> Cholesterol + fatty acids


Cholesterol + O2

---------------------------> 4-Cholesten-3- one + H2O

2

Peroxidase

2H2O

2 +Phenol+4-Aminoantipyrine --------------> - Red quinone + 4H

2O

REAGENT COMPOSITION

CHOLESTEROL R1 4 x 50 mL / 4 x 100 mL

Pipes buffer (pH 6.90) 50 mmol/L

Phenol 24 mmol/L

Sodium Cholate 0.5 mml/L

CHOLESTEROL R2 4 x 50 mL / 4 x 100 mL

Cholesterol esterase > 200 U/L

Cholesterol oxidase > 250 U/L

Peroxidase > 1000 U/L

4 – aminoantipyrine 0.5 mmol/L

CHOLESTEROL STANDARD 1 x 4mL

Cholesterol standard concentration 200 mg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-80C.

LINEARITY

This reagent is linear up to 500 mg/dL.

If the concentration is greater than linearity (500 mg/dL), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum / Plasma : 150 – 220 mg/dL


Dissolve contents of Reagent 2 (R2) with the amount of Reagent 1 (R1) indicatedon the vial label.


NOTE:Discard the working reagent if the blank absorbance exceeds 0.08.

PRECAUTION



CHOLESTEROL4 x 50 mL, 4 x 100 mL

11204002, 11204003

SAMPLE

Serum / Plasma (free of haemolysis).


Mode of Reaction End point


Wavelength I 505 (492 -540) nm

Wavelength II 630 nm

Temperature 370C

Standard Concentration 200 mg/dL

Blank Reagent

Linearity 500 mg/dL

Incubation time 5 min





Blank Standard Sample

Working Reagent 1000 µL 1000 µL 1000µL

Standard - 10 µL -

Sample - - 10 µL

Mix, and incubate for 5 min, at 370C. Measure the absorbance of sample and standardagainst reagent blank.

CALCULATION

Cholesterol Conc. (mg/dL) =

Absorbance of sample

----------------------------- x 200

Absorbance of standard

BIBLIOGRAPHY

1. Arntz, H. R.; Diagnostik der Hyperlipoproteinamein, Lab, Med. 3/177, (1979)2. Flegg, H.M.; Ann. Clin. Biochem, 10 (1973), 1350-13563. Siedel, J., Schlumberger, H., et al. ; J. Clin.Chem.Clin. Biochem.19 (1981), 838.4. Allain, C.C. et al.; Clin.Chem 20 (1974), 470


3

INTENDED USE:

This reagent is intended for in vitro quantitative determination of Cholinesterase inserum or plasma.

- Kinetic, Butyrylthiocholine method

- Linear up to 9084 U/L

- Stability of working reagent is 2 hours at 2-80C


Cholinesterase levels in serum are useful as a test of liver function, as an indicator ofpossible insecticide poisoning or for detection of patients with atypical forms ofenzyme.

Measurements of serum Cholinesterase activity can serve as sensitive measures of thesynthetic capacity of the liver if the patient’s normal level is known.

Elevated levels (marginal increase) are observed in patients with nephritic syndrome,thyrotoxicosis & haemochromatosis in obese, diabetic people & in patients withanxiety & other psychiatric states.

Decreased levels are found in patients with acute infections, pulmonary embolism &muscular dystrophy as well as after surgical procedures. After a myocardial infarction,the enzyme levels decreases until the 5th day & then begins a slow rise to normal.

Decreased levels are also found in chronic renal disease & in pregnancy.

PRINCIPLE

Cholinesterase

Butyrylthiocholine + H2o --------------->Thiocholine + Butyrate

Thiocholine + Dithio-bis-nitrobenzoate--------------> nitro -2- mercapto-5- benzoato

REAGENT COMPOSITION

CHOLINESTERASE R1 5 x 3 mL

Buffer Solution

Phosphate buffer, (pH 7.7) 50 mmol/L

CHOLINESTERASE R2 1 x 5 x 3 mL

Tablets

5,5 DTNB 0.25 mmol/L

Butyrylthiocholine 7 mmol/L


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C.

LINEARITY

This reagent is linear up to 9084 U/L.

If the concentration is greater than linearity (9084 U/L), dilute the sample with normalsaline and repeat the assay.Multiply the result with dilution factor.

NORMAL RANGE



Serum : 4659-14443 U/L


Dissolve one tablet R2 in one vial of Reagent 1 (R1). The stability of working reagentis 2 hours at 2-80c.

NOTE : Wait for 10 minutes for complete dissolution.

PRECAUTION



SAMPLE

Fresh plasma (free of haemolysis), serum.

CHOLINESTERASE5 x 3 mL

11205001


( OD/60 sec) ( OD/30 sec)

Mode of Reaction Kinetic Kinetic

Slope of reaction Increasing Increasing

Wavelength 405 nm 405 nm

Temperature 370C 370C

Factor 22710 45420

Linearity 9084 U/L 9084 U/L

Blank DI Water DI Water

Delay 2 sec 2 sec

No of reading 2 4

Interval 60 sec 30 sec

Sample volume 20 µL 20 µL

Reagent volume 3000 µL 3000 µL

Cuvette 1 cm light path 1 cm light path



Sample (dilute 1+1 with saline solution.) 20 µL

Mix and measure the change in absorbance per minute ( OD/60 sec)during 2 minutes. Or measure the change in absorbance per 30 sec ( OD/30 sec)during 2 minutes.

CALCULATION

Cholinesterase activity (U/L) =

( OD/60 sec) x 22710

or

( OD/30 sec) x 45420

BIBLIOGRAPHY

Kendel, M., Yotros.; Kin. Wschr. 45, 325 (1967)


4

INTENDED USE

This reagent is intended for in vitro quantitative determination of CK-MB in humanserum.

- Immuno – inhibition methodology


- Working Reagent Stable up to 8 days at 2- 80C

- Excellent pack sizes, Convenient for all laboratories


CK-MB is an enzyme formed by the association of two subunits from muscle(M) andnerve cells (B). CK-MB is usually present in serum at low concentration; it increasesafter an acute infarct of myocardium and later descends at normal levels. Also isincreased, rarely, in skeletal muscle damage.

Clinical diagnosis should not be made on a single test result; it should integrate clinicaland other laboratory data.

PRINCIPLE

An antibody to the anti CK-M inhibits completely CK-MM and subunit (M) of the CK-MB. The activity of the non – inhibited CK-B subunit is then assayed by the followingseries of reactions:

CK

Phosphocreatine + ADP ----> Creatine + ATP

HK

ATP + Glucose ----- > ADP + Glucose - 6- Phosphate

G6P- DH

G6P+ NADP+ ------------> 6 – Phosphogluconate + NADPH + H+

The rate of NADPH formation, measured photometrically, is proportional to thecatalytic concentration of CK –B present in the sample.

REAGENT COMPOSITION

CK-MB R1 5 x 2.5 mL

Buffer

Imidazol (pH 6.7) 100 mmol/L

Glucose 20 mmol/L

Magnesium acetate 10 mmol/L

EDTA 2mmol/L

CK-MB R2 5 x 2.5 mL

*Anti CK –M 200 U/L

ADP 2 mmol/L

AMP 5mmol/L

di-Adenosine-5-pentaphosphate 10 mmol/L

NADP+ 2 mmol/L

Hexokinase 2500 U/L

Glucose-6-phosphate dehydrogenase 1500 U/L

N-acetyl cysteine 20 mmol/L

Creatine Phosphate 30 mmol/L

*Anti CK-M sufficient to inhibit up to 2000 U/L of CK-MM

CK-MB Calibrator 1 x 2 mL

Calibrator concentration is mentioned on vial label


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C.

LINEARITY


If the concentration is greater than linearity (1000 U/L), dilute the sample with normalsaline and repeat the assay. Multiply result with dilution factor.

NORMAL VALUES

It is recommended that each laboratory should establish its own reference values.


250C 300 C 370C

CK-MB : > 10 U/L > 15 U/L >24 U/L


Reconstitute one tablet of R2 in one vial of R1.

The reconstituted reagent is stable for 8 days at 2-80C.

Calibrator: Reconstitute the calibrator with 2 mL distilled water and reconstitutedcalibrator is stable for:

24 hrs at 15 to 25oC

3 Days 2 to 8oC

1 Month at -15 to -25oC

PRECAUTION

To avoid contamination, use clean laboratory materials.


SAMPLE

Serum (Free of haemolysis)


OD/300 Sec OD/60 Sec

Mode of Reaction Fix Time Kinetic




Factor 1651 8254



Delay time 600 sec 180 sec

No of reading - 3

Delta time 300 sec 60 sec






Sample 40 µL

Mix and incubate at 370C for 10 minutes measure the initial absorbance(A1) and againtake the readings for 5 minutes(A2). Calculate the difference between absorbance A = (A2-A1), or mix and incubate at 370C for 3 minutes. Read the charge in absorbanceper minute ( OD/minutes) during 3 minutes.

CALCULATION

CK-MB Activity (U/L) = A x 1651

or

CK-MB Activity (U/L) = A/Min x 8254

BIBLIOGRAPHY

1. Abbot, B. et al.; Creatinine kinase. Kalpan, A. et al. Clin. Chem. The C.V.MosbyCo. St.Louis. Toronto Princeton 1984:1112 – 116

2. Gerhardt, W., et al.; Creatine K inase B-Subnit activity in serum afterimmunoinhibition of M-subunit activity.; Clin chem.1979; (25/7): 1274- 1280.

3. Young, D. S.; Effects of drugs on clinical Lab. Tests, 4th Ed AACC 2001.4. Burits, A. et al.; Tietz Textbook of Clinical Chemistry, 3rd Ed AACC 1999.5. Tietz, N. W. et al. Clinical Guide to Laboratory Tests, 3rd Ed AACC 1995.

CK MB with Calibrator 5 x 2.5 mL

11207004

5

INTENDED USE

This reagent is intended for in vitro quantitative determination of Creatine Kinase inhuman serum.

- Optimized IFCC Method


- Working Reagent is stable up to 5 days at 2-80C.

- Excellent pack Sizes, Convenient for all laboratories.


It is mainly found in all muscle (Cardiac & Skeletal) & brain tissues. It plays animportant role in energy storing mechanism of the tissues. It’s iso-enzymes: CK-MBmainly exists in cardiac muscle tissues, CK-MM in skeletal muscle tissues & CK-BBin brain.

Increased levels are found in myocardial infarction, muscular dystrophy,cerebrovascular-disease, pulmonary infarction, electrical shocks & hypothyroidisim.

Decreased levels are, some times seen in early pregnancy, alcohol, liver diseses, RA.

PRINCIPLE

Creatine kinase(CK) catalyses the reversible transfer of a phosphate group fromphosphocreatinine to ADP. This reaction is coupled to those catalyzed by hexokinase(HK) and glucose – 6- phosphate dehygrogenase (G6P-DH)

CK

Phosphocreatine + ADP ----- > Creatine +ATP

HK

ATP + Glucose ----- > ADP + Glucose – 6- phosphate

G6P – DH

G6P + NADP+ ------------- > 6- Phosphogluconate + NADPH + H+

The rate of NADPH formation, measured photometrically, is proportional to thecatalytic concentration of CK present in the sample.

REAGENT COMPOSITION

CREATINE KINASE R1 5 x 2.5 mL

Buffer

Imidazol pH 7.0 100 mmol/L

Glucose 20 mmol/L

Magnesium acetate 10 mmol/L

EDTA 2mmol/L

CREATINE KINASE R2 5 x 2.5 mL

Substrate

ADP 2 mmol/L

AMP 5 mmol/L

Di-Adenosine – 5-pentaphosphate 10mmol/L

NADP+ 2 mmol/L

Hexokinase 2500 U/L

Glucose -6-phosphate dehydrogenase 1500 U/L

N-acetyl cysteine 20 mmol/L

Creatine Phosphate 30mmol/L



LINEARITY

This reagent is linear up to 1000 U/L

If the concentration is greater than linearity (1000 U/L), dilute the sample withnormal saline and repeat the assay. Multiply the final result with dilution factor.

NORMAL VALUES

It is recommended that each laboratory establish its own reference value.


250C 300C 370C

Men up to : 80 U/L 130 U/L 195 U/L

Women up to : 70 U/L 110 U/L 170 U/L


Reconstitute one tablet R2 with one vial of R1 .

The reconstituted reagent is stable for 5 days at 2-80C.

PRECAUTION



CK-NAC5 x 2.5 mL

11206004

SAMPLE

Serum (Free of haemolysis)




Wavelength 340 nm

Temperature 370C

Factor 8095

Linearity 1000 U/L

Blank Distilled Water

Delay time 120 sec.

No of reading 3

Interval 60 sec






Sample 20 µL

Mix and incubate at 370C for 2 minutes, measure the change in absorbance perminute ( OD/min) during 3 minutes.

CALCULATION

Creatine Kinase Activity (U/L) = ( OD /min.) x 8095

BIBLIOGRAPHY

1. Abbot, B. et al. Creatinine kinase. Kalpan, A. et al.; Clin. Chem. The C.V.MosbyCo. St Louis. Toronto Princeton 1984:1112 – 116

2. Gerhardt, W. et al. Creatine K inase B-Subnit activity in serum afterimmunoinhibition of M-subunit activity; Clin. chem.1979; (25/7): 1274- 1280.

3. Young, D. S.; Effects of drugs on clinical Lab. Tests, 4th ed AACC 2001.4. Burits, A. et al. ; Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.6. Tietz, N. W. et al. Clinical Guide to Laboratory Tests, 3rd Ed AACC 1995.


6

INTENDED USE

This reagent is intended for in vitro quantitative determination of Glucose in serum,plasma & CSF.

- GOD –PAP methodology

- Linear upto 500 mg/dL

- Reconstituted stability 90 days at 2-80C

- No interferences from Bilirubin and Creatinine


Glucose is a major carbohydrate present in the blood & serves as a primary source ofenergy. It is usually obtained from ingested starch & sugar. The glucose concentrationis normally maintained at constant level. Excessive glucose is stored as inactiveglycogen mainly in the liver & little in the muscles.

Elevated blood glucose levels are found in diabetes mellitus, hyperthyroidism,hyperadrenalism & certain liver diseases.

Decreased levels are found in Insulinoma, hypothyroidism, hypopituitarism.

PRINCIPLE

Enzymatic colorimetric determination of glucose according to the following reaction.

Glucose Oxidase

Glucose+ O2

------------------> Gluconic acid + H2O

2

Peroxidase

2H2O

2+phenol + 4-Aminoantipyrine --------------- > red Quinonimine + 4H

2O

REAGENT COMPOSITION

GLUCOSE R1 4x100mL / 4x250mL / 4x500mL


Phenol 10 mmol/L

GLUCOSE R2 4x100mL / 4x250mL / 4x500mL

Glucose Oxidase > 10000 U/L

Peroxidase > 600U/L

4-Aminoantipyrine 270mmol/L

GLUCOSE STANDARD 1x4mL / 2x4mL

Glucose standard concentration 100 mg/dL

Note : For 4x500 mL and 4x250mL packs, Buffer will be provided separately



LINEARITY

This reagent is linear upto 500mg/dL

If the concentration is greater than linearity (500 mg/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum / Plasma : 70-105 mg/dL

C S F : 50 -70 mg/dL


Dissolve Reagent 2(R2 )with the volume of Reagent 1 (R1) indicated on the label.

The reconstituted reagent is stable for 90 days at 2-80c.

NOTE: Discard the working reagent if the blank absorbance exceeds 0.08.

PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / CSF.

GLUCOSE 4 x 100 mL,4 x 250 mL, 4 x 500 mL

11208001, 11208002, 11208003


Mode of Reaction Fixed time Kinetic


Wavelength 505 (500-540 nm)

Temperature 370C


Blank Reagent Blank

Linearity 500 mg/dL

Delay 30 sec.

Interval 60 sec




CALCULATION FOR FIXED TIME

Absorbance of Sample

Glucose conc (mg/dL)= ----------------------------------- x 100


GENERAL SYSTEM PARAMETER FOR ENDPOINT



Wavelength 505 nm

Temperature 370C


Blank Reagent Blank

Linearity 500 mg/dL







Working reagent 1000 µL 1000 µL 1000 µL

Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 10 minutes at 370C. Measure the change in absorbance ofstandard and sample against reagent blank.

CALCULATION FOR END POINT


Glucose Conc. (mg/dL) = ------------------------------ x 100


BIBLIOGRAPHY

Trinder, P.; Ann. Clin., Biochem. 6, (1969) 24


7

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGOT in serum orplasma.

- Proven IFCC methodology


- Reconstituted reagent stable up to 30 days at 2-80C


It is present in most of the tissues. Especially in cardiac muscle, liver cells, skeletalmuscle & kidneys. Injury to these tissues results in the release of the enzyme in bloodstream.

Increased levels are found in myocardial infarction. The duration & extent of increaseis related to the infract. GOT determination is of considerable value to differentiatemyocardial infraction from other cardiac disorders.

Increased levels are also found in various types of liver disease, skeletal muscle trauma& in renal diseases.

Decreased levels may be found in pregnancy, Beri-Beri & Diabetic ketoacidosis.

PRINCIPLE

Kinetic determination of Aspartate Aminotrasferase (AST) based upon the followingreaction.

AST/SGOT

L- Aspartate + alpha - ketoglutarate ------ > Oxaloacetate + L-Glutamate.

MDH

Oxaloacetate + NADH + H+ ------ > L- Malate + NAD+

AST : Aspartate aminotransferase.

MDH : Malate dehydrogenase.

REAGENT COMPOSITION

SGOT R1 2 x 53 mL / 4 x 50 mL

Tris Buffer (pH 7.8) 88 mmol/L

L-Aspartate 260 mmol/L

SGOT R2 5x 20 mL / 4x 50 mL

MDH > 600 U/L

LDH > 900 U/L

NADH 0.20 mmol/L

a -ketoglutarate 12 mmol/L

STORAGE & STABILITY


LINEARITY



REFERENCE RANGE



Females upto 31U/L at 37o CS

Males upto 35U/L at 37o C


Reconstitute the reagent 2 (R2 ) with the volume of reagent 1 (R1) mentioned on the

vial label. The reconstituted reagent is stable for 30 days at 2-80C.

NOTE: Discard the working reagent if the blank absorbance is less than 1.00 at 340nm.

PRECAUTION



SAMPLE


SGOT 5x 20 mL,4x 50 mL,

11213005, 11213003



Slope of reaction Decreasing

Wavelength 340 nm

Temperature 370C

Factor 1768

Linearity 350 U/L

Blank DI Water

Delay 60 sec.

No of reading 3

Interval 60 sec






Sample 100 µL

Mix and incubate at 370C for 1 minute. Measure the change in absorbance per minute( OD/min) during 3 minutes.

CALCULATION

SGOT activity (U/L) = ( OD/min) x 1768

BIBLIOGRAPHY

Expert panel on enzyme of the IFCC; Clin. Chim. Acta, 70(1976) F 19.


8

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGPT in serum orplasma.

- IFCC recommended methodology


- Reconstituted reagent stable up to 50 days at 2 - 80C


It is present in most of the tissues, but mainly found in the liver. Increased levels arefound in hepatitis, cirrhosis, obstructive jaundice & other hepatic disease. SGPT activityis markedly elevated even before clinical signs of jaundice become apparent in diseaseassociated with hepatic necrosis. Slight elevations are also found in myocardialinfraction.

PRINCIPLE

Kinetic determination of Alanine Aminotransferase (ALAT) according to the followingreaction.

ALT

L-Alanine + alpha-ketoglutarate ----- > Pyruvate +L-Glutamate

LDH

Pyruvate +NADH+ H+ ----- > L-Lactate +NAD+

ALT – Alanine aminotranferase

LDH - Lactate dehydrogenase

REAGENT COMPOSITION

SGPT R1 2 x 53 mL/ 4 x 50 mL

Tris buffer (pH 7.5) 110 mmol/L

L-Alanine 550 mmol/L

SGPT R2 5 x 20 mL/ 4 x 50 mL

LDH > 200 U/L

NADH 0.20 mmol/L

a-ketoglutarate 16 mmol/L



LINEARITY

This reagent is linear up to 350 U/L .


NORMAL RANGE



Serum up to : 49 U/L


Reconstitute the Reagent 2 (R2) with the volume of Reagent1 (R1) mentioned on thevial label. The reconstituted reagent is stable for 50 days at 2-80C.


PRECAUTION

To avoid contamination use clean laboratory wares.


SAMPLE


SGPT 5 x 20 mL, 4 x 50 mL

11214005,11214003




Wavelength 340 nm

Temperature 370C

Factor 1768

Blank DI Water

Linearity 350 U/L

Delay 60 sec.

No of reading 3

Interval 60 sec






Sample 100 µL


CALCULATION

SGPT activity (U/L) = ( OD/min) x 1768

BIBLIOGRAPHY

Expert panel on enzyme of the IFCC; Clin. Chim. Acta, 70 (1976) F19.


9

INTENDED USE

This reagent is intended for in vitro quantitative determination of triglycerides in serumor plasma.

- Proven GPO-POD methodology


- Extended stability, reconstituted reagent is stable up to 42 days at 2-80 C


Triglycerides are simple lipids, formed in the liver by glycerol & fatty acids. They aretransported by VLDL, LDL & constitute about 95% of fat, stored as source of energyin the tissue & plasma. Increased levels are found in hyperlipidemias, diabetes,nephrotic syndrome & hypothyroidism. Increased levels are risk factor forarteriosclerotic coronary disease, peripheral vascular disease, acute pancreatitis &hyperlipoproteinaemia. Decreased levels are found in malnutrition & hyperthyroidism.

PRINCIPLE

Enzymatic determination of triglyceride is based on the following reactions:

LPL

TGL+H2O ---- >Glycerol + Fatty acid

GK

Glycerol + ATP ---- > Glycerol-3-phosphate + ADP

Mg++

GPO

Glycerol-3-phosphate+O2------ > Dihydroxyacetone phosphate +H

2O

2

POD

2H2O

2+4-Aminoantipyrine+p-Chlorophenol ------ > Red Quinone+4H

2O

GPO = Glycereol-3-phosphate Oxidase

LPL = Lipoprotein Lipase

GK = Glycerol Kinase

REAGENT COMPOSITION

TRIGLYCERIDES R1 2 x 53 mL / 4 x 50 mL

Pipes –buffer (pH 7.00) 50 mmol/L

p-Chlorophenol 5.3 mmol/L

Magnesium salt 15 mmol/L

TRIGLYCERIDES R2 5 x 20 mL / 4 x 50 mL

Lipoprotein lipase >1100 U/L

Glycerol Kinase > 800 U/L

Glycerol -3-phosphate oxidase > 5000 U/L

Peroxidase 350U/L

4-Amino antipyrine 0.7 mmol/L / 0.3 mmol/L

TRIGLYCERIDES STANDARD 1 x 4 mL

Triglycerides standard concentration 200mg/dL

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Normal Value 65-165 mg/dL


Reconstitute the Reagent 2 (R2) with the volume of Reagent 1 (R1) indicated on thelabel.

Working reagent is stable for 42 days at 2-80C.

PRECAUTION



SAMPLE


TRIGLYCERIDES5 x 20 mL, 4 x 50 mL

11215002, 11215003


Mode of Reaction End Point


Wavelength I 505 nm (492-550 nm)


Temperature 370C


Linearity 1000 mg/dL

Blank Reagent







Working Reagent 1000 µL 1000 µL 1000 µL

Standard - 10 µL -

Sample - - 10 µL


CALCULATION


Triglycerides Con. (mg/dL) = ------------------------------ x 200 mg/dL


BIBLIOGRAPHY

1. Buccolo, G., David, M.; Clin.Chem., 19(1973) 4762. Wener, M., Gabrielson, D. G., Eastman, G.; Clin Chem 21 (1981) 2683. Annoni, G., Bottasso, B. M., Ciaci, D., Donato, M. F., Tripoli, A., Lamb, J.; J. Res.

Lab, Med., 9 (1982) 115


10

INTENDED USE

This reagent is intended for in vitro quantitative determination of urea in serum, plasma& urine.

- Modified Berthelot methodology


- Reconstituted reagent stability for 21 days at 2-80C.


Proteins cannot be stored in human body, so excess should be broken down. Aminoacids which from the components of proteins, break down to give ammonia. This istoxic & so through a series of chemical reactions (urea cycle) non toxic urea isproduced & this is released into the blood which is filtered in the kidney & excretedin the urine.

Elevated levels are seen during increased protein in breakdown, dehydration, vomiting,diarrhea. Also seen in any kind of renal disorder like Glomerular nephritis, chronicnephritis & Nephritic syndrome.

Decreased levels are found in liver failure & pregnancy

PRINCIPLE

Enzymatic determination of Urea according to the following reaction.

Urease

Urea + H2O ----------> 2NH

3 + CO

2

Nitroprusside

NH3 + salicylate ------------------> 2.2-Dicarboxy Indophenol

Hypochlorite

REAGENT COMPOSITION

UREA B COLOUR REAGENT R1 2 x 53 mL

Sodium Salicylate 80 mmol/L

Sodium Nitroprusside 4 mmol/L

Sodium Hypochlorite 45 mg /dL

UREA B R2 10 x 10 mL


Urease 20 KU/L

UREA B STANDARD 1 x 4 mL

Urea –B Standard concentration 40 mg/dL



LINEARITY



NORMAL RANGE



Serum : 10-55 mg/dL

Urine : 20-35 gm/24 hrs


Reconstitute 1 vial of reagent 2 (R2) with the volume of DI water as indicated on thevial label.


PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / Urine (1/100 Diluted)




Wavelength 600 nm (580-650 nm)

Temperature 370C


Linearity 200 mg/dL

Blank Reagent


Reagent volume 1000 µL + 1000 µL

Incubation time 5+5 min




Working reagent 1000 µL 1000 µL 1000 µL

Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 5 minutes at 370C then add.

Colour Reagent 1000 µL 1000 µL 1000 µL

Mix and incubate for 5 minutes at 370C then add.

DI Water 1000 µL 1000µL 1000 µL

Mix well & measure the absorbance of sample and the standard against the reagentblank.

CALCULATION


Urea Con. (mg/dL) = ---------------------------- X 40

Absorbance of Standard

BIBLIOGRAPHY

1. Wheatherburn M.W., Anal. Chem., 1967; 39;9712. Searcy, R.L., Reardon, J.E., Foreman, J.A.; Amer, J.Med, Tech., 1967; 33, 15-203. Henry, R. J.; Clin. Chem. Principle and Techniques, Harper & Row, New York,

1963.P.268

UREA - B 200 mL

11216001

11

INTENDED USE

This reagent is intended for in vitro quantitative determination of urea in serum,plasma & urine.

- Urease / GLDH methodology

- Linearity

Urea : 500 mg/dL

BUN : 235 mg/dL

- Working reagent stability is 42 days at 2-80C.


Proteins cannot be stored in human body, so excess should be broken down. Aminoacids which from the components of proteins, break down to give ammonia. Thisis toxic & so through a series of chemical reactions (urea cycle) non toxic urea isproduced & this is released into the blood which is filtered in the kidney & excretedin the urine.

Elevated levels are seen during increased protein breakdown, dehydration, vomiting,diarrhea. Also seen in any kind of renal disorder like Glomerular nephritis, Chronicnephritis & Nephritic syndrome.

Decreased levels are found in liver failure & pregnancy.

PRINCIPLE

Urease

Urea + H2O ---------- > 2NH

3 + CO

2

GLDH

2 NH3+2a- ketoglutarate + 2NADH --------- > 2L-Glutamate+2NAD+ + 2H

2O

REAGENT COMPOSITION

UREA U.V. R1 4 x 50 mL / 4 x 100 mL

Buffer (pH 7.55) 75 mmol/L

UREA U.V. R2 4 x 50 mL / 4 x 100 mL

ADP 0.66 mmol/L

GLDH >1000U/L

Urease > 30000 U/L

NADH 0.32 mmol/L

a-ketoglutarate 7.5 mmol/L

UREA U.V. STANDARD 1 x 4 mL

Standard concentration for Urea 50 mg/dL

Standard concentration for BUN 23.4 mg/dL

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Urea Urea – N

Sreum : 10-50 mg/dL 4.7 – 23 mg/dL

Urine /24 hr :20-36 gm/L 9.4-16.9 gm/L


Dissolve Reagent 2 (R2) with the volume of Reagent 1 (R1) indicated on the label.


Note: Discard the working reagent if the blank absorbance is less than 1.0 at 340 nm.

PRECAUTION



SAMPLE


Urine (1/100 diluted).

Do not use anticoagulants containing fluoride or ammonium ions.

UREA U.V 4 x 50 mL, 4 x 100 mL

11217001, 11217002


Mode of Reaction Fixed Time


Wavelength 340

Temperature 370C


Linearity 500 mg/dL

Blank DI Water

Delay time 30 sec

Interval 60 sec





Standard Sample

Working reagent 1000µL 1000 µL

Standard 10 µL -

Sample - 10 µL

Mix and read the optical density (T1) 30 seconds after the sample or standard

addition. Exactly 60 seconds after the first reading take second reading (T2).

CALCULATION

(T1 –T

2)of Sample

Urea Conc. (mg/dL) = ------------------------ x 50

(T1-T

2) of standard

(T1 –T

2)of Sample

Urea BUN Conc. (mg/dL) = ------------------------ x 23.4

(T1-T

2) of standard

BIBLIOGRAPHY

1. Talke, H. and Schubert, G. E.; Kiln-Wocchsr 19; 43: 1742. Thomas, L.; Labor and Diagnose, 1. Auflage, S. 346


12

INTENDED USE

This reagent is intended for invitro quantitative determination of Uric acid in serum,plasma & urine.

- Uricase - PAP methodology


- Reconstituted reagent stability up to 30 days at 2-80C

- Fast incubation, just 5 minutes at 370C


Uric acid is the end product of purine metabolism. Uric acid is excreted by the kidneys.Increased levels are found in gout, arthritis, impaired renal functions & starvation.Decreased levels are found in yellow atrophy of the liver, Wilson’s disease ,& Fanconissyndrom.

PRINCIPLE

Enzymatic determination of Uric acid according to the following reactions.

Uricase

Uric acid + 2H2O+O

2----------> Allantoin +CO

2 +H

2O

2Peroxidase

2H2O

2 + 4 -Aminoantipyrine +DHBS -------------> Red quinone + H

2O+ HCL

DHBS = 3,5-Dichloro -2-Hydroxybenzenesulfonic acid.

REAGENT COMPOSITION

URIC ACID R1 2 x 53 mL / 4 x 50 m L

Phosphate Buffer (pH 7.5) 100 mmol/L

DHBS 2 mmol/L

URIC ACID R2 5 x 20 mL / 4 x 50 mL

4 – Aminoantipyrine > 0.23mmol/L


Uricase > 60 U/L

URIC ACID STANDARD 1 x 4 mL

Uric acid standard concentration 6 mg/dL

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum / Plasma

Males : 3.4 - 7.0 mg/dL

Females : 2.5 - 6.0 mg/dL

Children : 2.5 - 5.5 mg/dL

Urine : 250 - 750 mg/ 24 hr urine


Reconstitute the reagent 2 (R2) with the volume of Reagent 1 (R1) indicated on thevial label.


PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / Urine (1/10 diluted with distilled water).

URIC ACID5x20 mL, 4 x 50 mL

11218002, 11218003




Wavelength I 505 nm (500 - 550)


Temperature 370C


Linearity 25 mg/dL

Blank Reagent



Incubation time 5 minutes





Standard - 20 µL -

Sample - - 20 µL

Mix and incubate 5 min. at 370C. Measure absorbance of sample and standard againstthe reagent blank.

CALCULATION


Uric Acid Con. (mg/dL) = ------------------------------ x 6


BIBLIOGRAPHY

1.Trivdi R.C., Revbar L., Berka, E., Strong L.,Clin. Chem. 24 (1978) 1908

2. Text Book of medical Laboratory Technology, sood


13

INTENDED USE

This reagent is intended for in vitro quantitative determination of Alkaline Phosphatasein serum or plasma.

- DGKC – SCE recommended procedure


- Working reagent can be prepared as per requirement

- Pack sizes to suit all types of laboratories


Alkaline phosphatase (ALP) is widely distributed throughout the body, but clinicallyimportant one for diagnostic reasons are in bone, liver, placenta & intestine. Growingbone is associated with the release of ALP and so in childhood the level of ALP isaround 3 times of that of adult. During pregnancy in 2nd & 3rd trimester the enzymerises considerably due to placenta releasing ALP. It can be used to examine placentalfunction.

Elevated levels are seen in bone diseases, e.g. pagets disease, rickets, osteoblasticmetastatic & in obstructive disease of biliary tract.


PRINCIPLE

Kinetic determination of ALP according to the following reaction

ALP

Para-nitrophenyl phosphate + H2O ----- > p-nitrophenol+Inorgnic

phosphate


REAGENT COMPOSITION

ALKALINE PHOSPHATASE(S.L) R1 2 x 8 mL /2 x 24 mL / 2 x 40 mL

Diethanolamine Buffer (pH 10.2) 125 mmol/L

Magnesium Chloride 0.625 mmol/L

ALKALINE PHOSPHATASE (S.L) R2 2 x 2 mL/2 x 6 mL / 2 x 10 mL

P-Nitrophenyl phosphate 50 mmol /L


The sealed reagents are stable upto the expiry date stated on the label, when storedat 2-80C.

LINEARITY

The reagent is linear, upto 700 U/L.


NORMAL RANGE


Women: 64 - 306 U/L

Men : 80 - 306 U/L

Children up to 15 yrs: < 644 U/L

Children up to 17 yrs: < 483 U/L


Mix 4 volume of Reagent 1 (R1) with 1 volume Reagent 2 (R2)


Note : Discard the working reagent if the blank absorbance exceeds 1.00 at 405 nm.

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure ofworking reagent to light.

SAMPLE


ALKALINE PHOSPHATASE (S.L) 2 x 10 mL, 2 x 30 mL, 2 x 50 mL

11401001, 11401005, 11401003




Wavelength 405 nm

Temperature 370C

Factor 2750

Blank DI water

Linearity 700 U/L

Delay time 60 sec

No of readings 3

Interval 60 sec






Sample 20 µL

Mix and incubate at 370C for one minute. Measure the change in absorbance perminute ( OD/min) during 3 minutes.

CALCULATION

ALP Activity (U/L) = ( OD / min.) x 2750

BIBLIOGRAPHY

1. Schlebusch, H., et al.; Dtsch .Med. Wschr. 99, 765 (1974)2. Z. Klin. Chem. Klin Biochem. 8,658 (1980) 10, 182 (1972)

ADL/V.02/JAN 2013

14

INTENDED USE

This reagent is intended for in vitro quantitative determination of amylase in serum,plasma & urine.

- CNPG3 methodology



Amylase occurs in the salivary glands, fallopian tubes & in pancreas.alpha-amylase is secreted by the pancreas from where it enters the duodenum, throughthe pancreatic duct. Any obstruction to these ducts causes alpha-amylase enzyme toenter the blood stream.

Elevated levels seen in acute pancreatitis, peptic ulcers, biliary disease, parotitis & otherintestinal obstructions.

Decreased levels are seen in chronic pancreatic disorders having pancreatic celldestruction.

PRINCIPLE

Amylase

5CNPG3 ------------> 3 CNP +2CNPG2+3 Maltotriose + 2 Glucose.

CNP = 2-Chloro-4-nitrophenol

CNP-G2= 2-chloro -4-nitrophenyl-a-maltoside

REAGENT COMPOSITION

ALPHA AMYLASE (S.L) R14x 5 mL / 4 x 10 mL

MES Buffer (pH6.0) 50 mmol/L

CNPG3

2.27 mmol/L

Calcium chloride 60 mmol/L

Sodium chloride 70 mmol/L

Activator 900 mmol/L



LINEARITY

The reagent is linear, up to 2000 U/L.


NORMAL RANGE


Serum / plasma : 25 - 86 U/L

Urine : < 470 U/L

PREPARATION AND STABILITY OF REAGENT

The reagent is ready to use.

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure of reagentto light.

This reagent is very sensitive to external contamination, ie Saliva, Sweat etc whichcontains alpha-amylase. Handle with gloves & keep vial tightly sealed after use.

Discard reagent if it turns cloudy.

SAMPLE

Fresh serum / plasma (free of haemolysis) / Urine (1/3 diluted)




Wavelength 405 nm

Temperature 370C

Factor 3178

Linearity 2000 U/L

Blank DI water

Delay time 60 sec

No.of readings 3

Interval 60 sec





Reagent (R1) 1000 µL

Sample 25 µL

Mix and incubate for 1 min. at 370C . Measure the change in absorbance per minute( OD/min) during 3 minutes.

CALCULATION

alpha-Amylase activity (U/L) = ( OD/ min.) x 3178

BIBLIOGRAPHY

1. Junge, W., et al.; Clin. Biochem. 22, 109(1989)2. Hohenwallnern, W.; J.Clin. chem. Clin. Biochem. 27,97(1989)

4 x 5 mL, 4 x 10 mL

11402001, 11402002AMYLASE (S.L)

15

INTENDED USE

This reagent is intended for in vitro quantitative determination of Cholesterol in serumor plasma.

- CHOD-PAP methodology


- Contains LCF (Lipaemic clearing factor) which minimizes rerun


It is the main lipid found in the blood, bile & brain tissues. It is also one of the mostimportant steroids of the body & is a precursor of many steroid hormones. Two thirdsof cholesterol present in the blood is esterified. The liver metabolizes the cholesterol& it is transported in the blood stream by lipoproteins.

Increased levels are found in hypercholesterolaemia, hyperlipidaemia, hypothyroidism,uncontrolled diabetes, nephritic syndrome & cirrhosis.

Decreased levels are found in malabsorption, malnutrition, hyperthyroidism, anaemia& liver diseases.

PRINCIPLE

Enzymatic colorimetric determination of total cholesterol according to the followingreactions.


Cholesterol ester +H2O --------------------------> Cholesterol + fatty acids

Cholesterol Oxidase

Cholesterol + O2

--------------------------> 4-Cholesten-3- one + H2O

2

Peroxidase

2H2O

2 +Phenol+4-Aminoantipyrine --------------> - Red quinone + 4H

2O

REAGENT COMPOSITION

CHOLESTEROL (S.L) R1 5 x 25 mL / 5 x 100 mL / 4 x 250 mL


Phenol 24 mmol/L

Sodium Cholate 0.5 mmol/L

4-aminoantipyrine 0.5 mmol/L

Cholesterol Esterase > 180 U/L

Cholesterol Oxidase > 200 U/L


CHOLESTEROL STANDARD 1 x 4 mL

Cholesterol std. concentration 200 mg/dL


The sealed reagents are stable upto the expiry date stated on the label when stored at2- 80C.

LINEARITY

This reagent is linear upto 600 mg/dL.


NORMAL RANGE



Serum / Plasma : 150 – 220 mg/dL


The reagent is ready to use.

PRECAUTION


Avoid direct exposure of reagent to light.

SAMPLE


CHOLESTEROL (S.L)5 x 25 mL, 5 x 100 mL, 4 x 250 mL

11403002, 11403003, 11403007




Wavelength I 505 (492 -550) nm


Temperature 370C


Blank Reagent

Linearity 600 mg/dL








Standard - 10 µL -

Sample - - 10 µL

Mix,and incubate for 5 min.at 370C. Measure the absorbance of sample and standardagainst reagent blank.

CALCULATION

Cholesterol Conc. (mg/dL) = Absorbance of sample x 200


BIBLIOGRAPHY

Allain, C.C., et al.; Clin.Chem 20 (1974), 470

ADL/V.02/JAN 2013

16

INTENDED USE

This reagent is intended for in vitro quantitative determination of cholinesterase inserum or plasma.

- New DGKC method

- Ready to use liquid stable reagents

- High linearity and high sensitivity


Cholinesterase also known as acetyl-cholinesterase and is found mainly in the nerveendings and the grey matter of brain. It hydrolyses acetylcholine, released at the nerveendings to mediate transmission of impulses. A similar enzyme acyl cholineacylhydrolase also known as pseudocholinesterase is present in the serum, brain,heart, liver and pancreas but its biological role is unknown.

Cholinesterase levels in serum are useful as a test of liver function and as an indicatorof possible insecticide poisoning. Among the organic phosphorous compounds thatinhibit cholinesterase activity are mainly insectisides such as Parathion, Sarin andTetraethylpyrophosphate. During poisoning the level of enzymes decreases as itsactivity is inhibited.

PRINCIPLE

Cholinesterase catalyses the hydrolysis of butyrylthiocholine substrate formingbutyrate and thiocholine.

CHE

butyrylthiocholine+H2O`------- > Thiocholine + Butyrate

Thiocholine reduces hexacyanoferrate(3) to hexacyanoferrate(2)

Thiocholine + hexacyanoferrate(3)---------> Hexacyanoferrate(2)

The decrease of absorbance is followed at 405 nm and is proportional to the activityof cholinesterase in the sample.

REAGENT COMPOSITION

CHOLINESTERASE(S.L) R1 2 x 8 mL

Pyrophosphate buffer, (pH 7.6) 75 mmol/L

Hexa cyanoferrate(3) 2 mmol/L

CHOLINESTERASE (S.L)R2 2 x 2 mL

Butyrylthiocholine 15 mmol / L


The sealed reagents are stable upto the expiry date stated on the label, when storedat 2- 80C.

LINEARITY

This reagent is linear upto 41400 U/L.

The lower detection limit is 50 U/L.

NORMAL RANGE



Females : 3930 – 10800 U/L

Males : 4620 – 11500 U/L


Mix 4 parts of R1 with one part of R2 to prepare working reagent.

The working reagent is stable only for 3 hours at 15 -250C.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent bottles immediately after use. Avoid direct exposureof working reagent to light.

SAMPLE

Use fresh serum / plasma.

Do not use sodium fluoride as an anticoagulant because it inhibits cholinesterase.Samples are stable for 15 days at 2-80C.

CHOLINESTERASE(S.L) 2 x 10 mL

11419002




Wavelength 405 nm

Temperature 370C

Factor 22653

Linearity 41400 U/L

Blank DI Water

Delay 30 sec

No of reading 3

Interval 30 sec






Sample 50 µL

Mix and incubate at 370C for 30 seconds . Measure the change in absorbance per30 seconds during 90 seconds.

CALCULATION

Cholinesterase activity (U/L) = ( OD/ 30 sec) x 22653

Unit conversion : U/L x 0.01667 = µ katal/L

INTERFERENCES

Test will not be affected by

Bilirubin upto20 mg/dL

Hemoglobin upto500 mg/dL

Triglycerides upto1000 mg/dL

BIBLIOGRAPHY

1. Eur. J. Clin. Chem; Clin. Biochem 30, 163(1992)2. Tietz, Text book of Clinical chemistry, 2nd Edition, Brutis, Ashwood (1994)3. Knedel, M and Bottger, R; Klin. Wschr. 1967; 45;30


17

INTENDED USE

This reagent is intended for in vitro quantitative determination of CK-MB in humanserum or plasma.

- Immuno – inhibition methodology


- Convenient pack sizes for all laboratories


CK – MB levels increases significantly 4-6 hours following a myocardial infarction &peak at around 12 to 24 hours after the infarct. The levels return to normal in caseof no further myocardial damage after 24 - 48 hours. Hence the increased levels ofCK-MB along with elevated levels of CK-NAC is a good indicator of myocardial infarction.

PRINCIPLE

The procedure involves measurement of CK-activity in the presence of an antibodyto CK-M monomer. This antibody completely inhibits the activity of CK-MM & halfof the activity of CK-MB, while not affecting the B subunit activity of CK-MB &CK –BB. Then we use CK method to quantitatively determine CK-B activity. The CK-MBactivity is obtained by multiplying the CK-B activity by two.

REAGENT COMPOSITION

CK-MB (S.L) R1 2 x 10mL / 2 x 16 mL

Imidazole (pH 6.7) 125 mmol/L

D-Glucose 25 mmol/L

N-Acetyl –L- Cysteine 25 mmol/L

Magnesium acetate 12.5 mmol/L

NADP 2.52 mmol/L

EDTA 2.02 mmol/L

Hexokinase > 6800 U/L

Anti human polyclonal CK-M antibody (sheep) sufficient to inhibit up to 2000 U/Lof CK-MM.

CK-MB (S.L) R2 2 x 2.5 mL / 2 x 4 mL

Creatine phosphate 250 mmol/L

ADP 15.2mmol/L

AMP 25 mmol/L

Diadenosine pentaphosphate 103 mmol/L

G-6-PDH > 8800 U/L



LINEARITY

This reagent is linear upto 600 U/L.


NORMAL VALUES




% CK- MB : 6 - 25 %


Mix 4 volume of Reagent 1 (R1) with 1 volume of Reagent 2 (R2).


NOTE: Discard the working reagent if the blank absorbance exceeds 1.0 at 340 nm.

PRECAUTION



SAMPLE

Serum / plasma (Free of haemolysis)

CK-MB (S.L) 2 x 12.5 mL, 2 x 20 mL

11405007, 11405002




Wavelength 340 nm

Temperature 370C

Factor 8254

Linearity 600 U/L

Blank DI Water

Delay 100 sec

No of reading 5

Interval 60 sec






Sample 40 µL

Mix and incubate at 370C for100 seconds. Read the change in absorbance per minute( OD/ minute) during 5 minutes.

CALCULATION

CK-MB Activity (U/L) = OD/Min x 8254

BIBLIOGRAPHY

1. DGKC, J.Clin. Chem Clin. Bioch. 15, 255(1977)2. Di. Witt, C Trendelendurg, J. Clin. Chem, Clin Bioch. 20, 235 (1982)


18

INTENDED USE

This reagent is intended for in vitro quantitative determination of Creatine Kinase inhuman serum or plasma.

- Optimized IFCC Method


- Working Reagent can be prepared as per requirements


It is mainly found in all muscle (Cardiac & Skeletal) & brain tissues. It plays an importantrole in energy storing mechanism of the tissues. It’s iso-enzymes: CK-MB mainly existsin cardic muscle tissues, CK-MM in skeletal muscle tissues & CK-BB in brain.

Increased levels are found in myocardical infarction, muscular dystrophy,cerebrovascular-disease, pulmonary infarction, electrical shocks & hypothyroidisim.

Decreased levels are, some times seen in early pregnancy, alcoholic liver diseases andRA.

PRINCIPLE

Kinetic determination of Creatine Kianse is based on following reactions:-

CK

Creatine Phosphate + ADP ---- > Creatine +ATP

HK

ATP + Glucose ---- > ADP + Glucose – 6- phosphate

G6P – DH

G-6-P + NADP+ ----------- > D-Gluconate -6-phosphate + NADPH + H+

CK – Creatine Kinase

HK – Hexokinase

G-6-P-Glucose-6-phosphate

G-6-PDH-Glucose-6-Phosphate dehydrogenase.

REAGENT COMPOSITION

CK-NAC (S.L) R1 2 x 8 mL / 2 x24 mL

Immidazole buffer 125 mmol/L

D-Glucose 25 mmol/L

N-Acetyl-L-cysteine 25 mmol/L


NADP 2.4 mmol/L

EDTA 2.0 mmol/L

Hexokinase >6800 U/L

CK-NAC (S.L) R2 2 x 2 mL / 2 x 6 mL




LINEARITY

This reagent is linear up to 1700 U/L


NORMAL VALUES



Men upto : < 171 U/L

Women upto : < 145 U/L


Mix 4 volume of Reagent 1 (R1) with 1 volume of Reagent 2 (R2)


NOTE: Discard the working reagent if the blank absorbance exceeds 1.0 at 340 nm.

PRECAUTION



SAMPLE

Serum / Plasma (Free of haemolysis)

CK-NAC (S.L) 2 x 10 mL, 2 x 30 mL

11404001,11404005




Wavelength 340 nm

Temperature 370C

Factor 4127

Linearity 1700 U/L

Blank Distilled Water

Delay time 100 sec.

No of reading 3

Interval 60 sec






Sample 40 µL

Mix and incubate at 370C for 100 seconds. Read the change in absorbance per minute( OD/min) during 3 minutes.

TWO REAGENT PROCEDURE

Reagent 1 R1 – 200 µL

Reagent 2 R2 – 50 µL

Mix and wait for 25 seconds, add sample 10 µL, mix well and incubate for 2 minutesat 370C, measure the variation of absorbance per minute during 3 minutes.

CALCULATION

Creatine Kinase Activity (U/L) = ( OD /min.) x 4127

BIBLIOGRAPHY

1. DGKC, J.Clin. chem.Clin. Bioch.15, 255 (1977)

2. Di. Witt, C. Tren delenburg, J. Clin chemie, Clin. Bioch. 20,235 (1982)


19

INTENDED USE

This reagent is intended for in vitro quantitative determination creatinine in serum orurine.

- High Linearity of 200 mg/dL

- No sample dilution

- Ready to use reagents


Creatinine is formed in muscles from phosphocreatinine. It is an important form ofenergy by being a store of high energy phosphate. Creatinine determinations have oneadvantage over urea determination that it is not affected by a high protein diet.

Serum creatinine is more specific & sensitive indicator of renal function. Simultaneousestimations of serum urea & creatinine provides better information. Serum ureanitrogen & creatinine ratio is > 15 in prerenal failure & < 10 in renal failure. Decreasedlevels are found in muscle dystrophy.

PRINCIPLE

Creatininase

Creatinine +H2O ------------------> Creatine

Creatinase

Creatine + H2O ------------------> Sarcosine +Urea

Sarcosine Oxidase

Sarcosine + O2+H

2O ------------------------> Glycine + HCHO + H

2O

2

Peroxidase

2 H2O

2+ 4-AA *1+ TOOS *2 ---------------> Quinone pigment +4 H

2O

* 1 : 4- Aminoantipyrine,

* 2 : N-ethyl-N-(2-hydroxy -3-sulfopropyl)-m-toluidine

Creatinine concentration can be obtained by measuring quinone pigmentphotometrically.

REAGENT COMPOSITION

CREATININE (S.L) R1 2 x 30 mL / 2 x 45 mL / 4x 45 mL

Creatinase 175000 IU/L

Sarcosine Oxidase 1500 IU/L

TOOS 1.13 mmol

CREATININE (S.L) R2 2 x 10 mL / 1 x 30 mL / 2 x 30 mL

Creatininase 75000 IU/L

Peroxidase 4500 units/L

4-AA 0.75 mmol

CREATININE STANDARD 1 x 4 mL

Creatinine Std. Conc. 2 mg/dL

STORAGE & STABILITY


LINEARITY

This reagent is linear upto 200 mg/dL.

If the concentration is greater than linearity dilute the sample with normal saline andrepeat the assay. Multiply the result with dilution factor.

REFERENCE VALUES



Serum : male : 0.6 – 1.1 mg/dL

: Female : 0.5- 0.8 mg/dL

Urine : Male : 1070-2150 mg/dL (24 hrs accumulated urine)

: Female : 769 – 1200 mg/dL (24 hrs accumulated urine)


Reagent 1 & Reagent 2 are ready to use.

PRECAUTION



SAMPLE

Fresh Serum/urine(24 hour)

ENZYMATIC CREATININE2 x 40 mL, 2 x60 mL, 4 x 60 mL

11420002, 11420003, 11420004




Wavelength I 546 nm

Wavelength II 630

Temperature 370C

Standard Conc. 2 mg/dL

Linearity 200 mg/dL

Blank Reagent

Incubation time 10 min.


Reagent 1 volume 450 µL




Blank Calibrator Sample

Reagent 1 450 µL 450 µL 450 µL

Calib./Std - 10 µL -

Sample - - 10 µL

Mix and incubate for 5 minutes at 370C then add,


Mix and incubate for 5 minutes at 370C. Measure the absorbance of sample and thestandard against the reagent blank.

CALCULATION


Creatinine Conc (mg/dL) = -------------------------------- x standard conc.


BIBLIOGRAPHY

Artiss, J.D., Mc Enroe, R.J., Zak, B.; Clin.Chem, 30 (1984)1389.


20

INTENDED USE

This reagent is intended for in vitro Quantitative determination of gamma GT in serum.

- Szasz methodology


- Working reagent can be prepared as per requirements


GGT activity is elevated in all forms of liver diseases. It is highest in cases of intrahepaticor post hepatic biliary obstruction. (It may be 5 to 30 times higher than normal). It ismore sensitive than alkaline phosphatase NTP leucine amino peptidase andtransaminases in detection of obstructive jaundice, cholangitis, cholecystis neoplasm,it rises earlier than other enzyme and persists longer.

Moderate increase is observed in infectious hepatitis (2 to 5 times). Increases mayalso be observed in cases of drug intoxication, acute and chronic pancreatitis.

PRINCIPLE

Kinetic determination of Gamma GT according to the following reaction.

Gamma GT

GLUPA-C+Glycylglycine ---------------> L-Gamma-Glutamyl-GlycyLglycine +

5-Amino-2-nitrobenzoic acid.

GLUPA-C: L-Gamma -Glutamyl-3 Carboxy-p-nitroanilide

REAGENT COMPOSITION

Gamma GT R1 2 x 8 mL / 2 x 24 mL

Tris buffer pH (8.25) 133 mmol/L

Glycylglycine 138 mmol/L

Gamma GT R2 2 x 2 mL / 2 x 6 mL

GLUPA-C 23 mmol/L



LINEARITY


If the concentration is greater than linearity (232 U/L) dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL VALUES


The following values may be used as guide line.

Female : 5 - 32 U/L

Male : 10 - 45 U/L


Mix 4 volume of Reagent 1 (R1) with 1 volume of reagent 2 (R2). This working reagentis stable for 21 days at 2-80C.

Note: Discard the working reagent if the blank absorbance exceeds 1.0 at 405 nm.

PRECAUTION:To avoid contamination, use clean laboratory wares.Avoid direct exposure of working reagent to light.

SAMPLE

Fresh Serum (free of haemolysis).




Wavelength 405 nm

Temperature 370C

Factor 1158

Linearity 232 U/L

Blank DI Water

Delay 60 sec

No of readings 3

Interval 60 sec






Sample 100 µL

Mix and incubate for 1 minute at 370C. Read the change in absorbance per minute( OD/min) during 3 minutes.

CALCULATION

Gamma GT activity (U/L) = ( OD/min) x 1158

BIBLIOGRAPHY

1. Szasz, G.; Clin.Chem., 22, (1976),2051.2. Scand. J.Clin. Lab.Invest 36:711 (1976)3. Tietz, N.W.; Text book of Clin.Chem. 678-686 : (1986)

GAMMA GT(S.L) 2 x 10 mL, 2 x 30 mL

11416001, 11416005

21

INTENDED USE

This reagent is intended for in vitro quantitative determination of Glucose in serum,plasma & CSF.

- GOD-PAP methodology



Glucose is a major carbohydrate present in the blood & serves as a primary source ofenergy. It is usually obtained from ingested starch & sugar. The glucose concentrationis normally maintained at a constant level. Excessive glucose is stored as inactiveglycogen mainly in the liver & little in the muscles.



PRINCIPLE


Glucose Oxidase

Glucose+ O2 + H

2O -----------------------> Gluconic acid + H

2O

2

Peroxidase

2H2O

2+phenol + 4-Aminoantipyrine -------------------> Quinonimine + 4H

2O

REAGENT COMPOSITION

GLUCOSE (S.L) R1 5 x 100 mL / 1 x 1000 mL

Tris Buffer, (pH 7.40) 92 mmol/L

Phenol 0.3 mmol/L

Glucose Oxidase 15000 U/L

4- Aminophenazone 2.6 mmol/L

GLUCOSE STANDARD 1 x 4 mL

Glucose standard concentration 100 mg/dL



LINEARITY

This reagent is linear up to 600 mg/dL


NORMAL RANGE



Serum / Plasma : 70-105 mg/dL

C S F : 50 -70 mg/dL


The Reagent is ready to use.

PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / CSF




Wavelength 505 (490-550 nm)

Temperature 370C


Linearity 600 mg/dL

Incubation Time 10 Minutes

Blank Reagent







Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 10 minutes at 370C. Read the absorbance of standard and sampleagainst reagent blank.

CALCULATION


Glucose Conc. (mg/dL) = -------------------------- x 100


BIBLIOGRAPHY

1. Trinder, P.; Ann Clin Biochem. 6,24 (1969)2. Dingeon, B.; Ann.Bio.Clin 33,3 (1975)3. Lott, J. ; Clin.Chem. 21, 1754 (1975)

GLUCOSE (S.L)5 x 100 mL, 1 x 1000 mL

11406001, 11406002

22

INTENDED USE

This reagent is intended for in vitro quantitative determination of HDL Cholesterol inserum

- Direct determination of HDL Cholesterol

- Enzyme Selective Protection Method




Blood total cholesterol levels have long been known to be related to coronary heartdisease (CHD). In recent years, in addition to total cholesterol, high densitylipoprotein cholesterol (HDL-C) has become an important tool used to assess anindividual risk of developing CHD since a strong negative relationship betweenHDL - C concentration and the incidence of CHD was reported.

PRINCIPLE

The reaction between cholesterol other than HDL and the enzyme for cholesterolassay is suppressed by the electrostatic interaction between polyanions & cationicsubstances. Hydrogen peroxide is formed by the free cholesterol in HDL by cholesteroloxidase. Oxidative condensation of EMSE and 4-AA is caused by hydrogen peroxidein the presence of peroxidase, and the absorbance of the resulting red-purple quinoneis measured to obtain the cholesterol value in HDL

Polyanions

Other lipoproteins than HDL --------------> Suppress reaction with enzyme

Cationic substances

cholesterol esterase

HDL (cholesterol esters) + H2O --------------------------> HDL (free cholesterol) +

Free fatty acids

cholesterol oxidase

HDL (free cholesterol) + O2

+ H+ -------------------------> Cholestenone + H2O

2

Peroxidase

2H2O

2 + 4-AA + EMSE + H

3 + O ---------------> Red-purple quinone + 5H

2O

REAGENT COMPOSITION

HDL –C DIRECT R1 2 x 30 mL / 2 x 45 / 4 x 45 mL

N—ethyl-N-(3-methylphenyl)-N’succinylethyenediame(EMSE)

HDL –C DIRECT R2 2 x 10 / 1 x 30 / 2 x 30 mL

Cholesterol Oxidase

4-Aminoantipyrin(4-AA)

HDL –C DIRECT CALIBRATOR 1 x 3 mL


The sealed reagents are stable upto the expiry date stated on the label, when storedat 2 - 80C, protected from light. Do not freeze.

LINEARITY

This reagent is linear upto 150 mg/dL

If the concentration is greater than linearity (150 mg/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor

NORMAL RANGE



Male : 35 - 80 mg/dL

Female : 42 - 88 mg/dL


The Reagent1 & Reagent 2 are ready to use.

Calibrator : Reconstitute with 3 mL of distilled water. Let it stand for 30 minutes atroom temperature. Dissolve the content of the vial by swirling gently to avoid theformation of foam.

Stability : Reconstituted calibrator is stable only for 7 days at 2- 80C.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light. Do not blow into the reagent bottles.

SAMPLE

Fresh serum (free of haemolysis)




Wavelength I 578 nm(578-610)

Wavelength II 630 nm (630-700)

Temperature 370C

Calibrator Con As on the vial

Linearity 150 mg/dL

Incubation time 5 min+5 min

Blank Reagent

Sample volume 5 µL

Reagent volume 450 µL+150 µL




Reagent1 450 µL 450 µL 450 µL

Calibrator - 5 µL -

Sample - - 5 µL

Mix and incubate for 5 minutes at 370C.


Mix and incubate for 5 minutes at 370C. Measure the absorbance of calibrator &sample against reagent blank at 578nm/ 630 nm.

CALCULATION


HDL-C Concentration (mg/dL) = ------------------------------ x Calibrator Concentration

Absorbance of Calibrator

ALTERNATIVE PROCEDURE

Mode of Reaction Differential


Wavelength I 600 nm


Temperature 370C

Calibrator Concentration As on the vial label

Linearity 150 mg/dL

Blank Reagent

Incubation time 5 minutes+5 minutes

Sample volume 3 µL





Reagent1 270 µL 270 µL 270 µL


Sample - - 3 µL

Mix and incubate for 5 minutes at 370C. Measure the absorbance (OD1) at 600 nm/700 nm.



CALCULATION

(OD2-OD1) Sample

HDL-C Concentration (mg/dL) = ---------------------------- x Calibrator Conc.

(OD2-OD1) Calibrator

INTERFERING SUBSTANCES

Test will not be affected by:

Bilirubin up to 40 mg/dL

Ascorbic acid up to 50 mg/dL

Hemoglobin up to 500 mg/dL

Triglyceride up to 1000 mg/dL

*(when triglyceride in a sample exceeds 1000 mg/dL, dilute the sample 1+9 withsaline, repeat the assay and multiply result by 10)

BIBLIOGRAPHY

1. Williams, P., et al.; High density lipoprotein and coronary risk factor, Lancet. 1:72(1979)

2. Gordon,T., Castelli, W.P., Hjortland, M.C. et al. Am. J.Med. 62, 707-714 (1977)3. Rifai,N.and Warnick, G.R.,Ed. Laboratory Measurement of Lipids, Lipoproteins

and Apolipoproteins AACC Press. Washington, DC,USA,1994

HDL CHOLESTEROL (D) WITH CALIBRATOR 2 x 40 mL / 2 x 60 mL / 4 x 60 mL

11414003, 11414005, 11414004


23

INTENDED USE

This reagent is intended for in vitro quantitative determination of Lactatedehydrogenase in serum or plasma.

- Based on SCE recommended method



This enzyme is found in all organ cells, but especially plentiful in cardiac & skeletalmuscle, liver, kidney & RBC. LDH is found in the form of iso-enzymes based on theirelectrophoretic mobility with each iso-enzymes being primarily from different organs.

Elevated levels are found in myocardial infarction, liver diseases, hemolytic anaemias,pernicious anaemia, Leukemia & Pulmonary diseases. Elevations in acute MI reachesa peak in 48-72 hrs. belonged elevations, (10-14 days) are useful in the late diagnosisof the condition.

PRINCIPLE

Kinetic determination of lactate dehydrogenase according to the following reaction.

LDH

Pyruvate + NADH + H+ --------------> L-Lactate +NAD+

REAGENT COMPOSITION

LDH-P (S.L) R1 2 x 8mL / 2 x 24mL


Pyruvate 1.6 mmol/L


LDH-P (S.L) R2 2 x 2 mL / 2 x 6 mL

NADH 240 mmol/L

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum /Plasma : 225-450 U/L


Mix 4 volumes of Reagent 1 (R1) with 1 volume of Reagent 2 (R2)



PRECAUTION



SAMPLE





Wavelength 340 nm

Temperature 370C

Factor 16030

Linearity 2400 U/L

Delay 60 sec.

No of reading 3

Interval 60 sec






Sample 10 µL


CALCULATION

LDH-P activity (U/L) = ( OD/min) x 16030

BIBLIOGRAPHY

1. Z.Klin. chem. Klin Biochem.8,658 (1970), 1, 1820(1972)2. Wei Bhaar, D., et al.; Med.Welt 26,387 (1975)

LDH-P (S.L)2 x 10 mL, 2 x 30 mL

11407001, 11407004

24

INTENDED USE

This reagent is intended for in vitro quantitative determination of LDL Cholesterol inserum or plasma

- Direct determination of LDL Cholesterol

- Enzyme selective protection method




Blood total cholesterol levels have long been known to be related to coronary heartdisease (CHD). In recent years, in addition to total cholesterol, low densitylipoprotein cholesterol (LDL-C) has become an important tool used to assess anindividual risk of developing CHD since a strong positive relationship between LDL-C concentration and the incidence of CHD was reported. LDL Cholestrol acts as akey factor in the pathogenesis of atherosclerosis and coronary artery disease.

PRINCIPLE

The LDL-C Direct is a homogeneous assay for directly measuring LDL-C levels inserum or plasma without the need for any off-line pretreatment or centrifugation

In the first reaction, non LDL unesterified cholesterol is subject to an enzymereaction and the peroxide generated is consumed by peroxidase in the presence of4-AAP to yield a colorless product. The second reagent consists of a detergentcapable of solubilizing LDL specifically. Cholesterol esterase and chromogeniccoupler react with this solubilize LDL-C to develop color. The intensity of colorformed is directly proportional to the concentration of LDL-C.

REAGENT COMPOSITION

LDL –C DIRECT R1 2 x 30 mL / 2 x 45 mL

4-Aminoantipyrin 0.5 mmol/L

CHE

CO 1.2 U/mL

Peroxidase

Good’s buffer pH 6.3

LDL –C DIRECT R2 2 x 10 / 1 x 30 mL

N,N-bis(4-sulfobutyl)-m-toluidine

disodium salt (DSBmT) 1.0 mmol/L


LDL –C DIRECT CALIBRATOR 1 x 3 mL


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C, protected from light. Do not freeze.

LINEARITY


If the concentration is greater than linearity (450 mg/dL), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor

NORMAL RANGE



Desirable < 130 mg/dL

Borderline High Risk for CHD 130-159 mg/dL

High Risk for CHD >160 mg/dL.


The Reagent 1 & Reagent 2 are ready to use.

Calibrator : Reconstitute with 3 mL of distilled water. Let it stand for 2 hours at roomtemperature. Dissolve the content of the vial by swirling gently to avoid theformation of foam.

Stability : Reconstituted calibrator is stable only for 7 days at 2- 80C.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light. Do not blow into the reagent bottles.

SAMPLE

Fresh serum (free of haemolysis) / EDTA Plasma


Mode of Reaction Differential


Wavelength I 546 nm


Temperature 370C


Linearity 450 mg/dL

Blank Reagent

Incubation time 5 min + 5 min

Sample volume 3 µL




DIFFERENTIAL MEASUREMENT




Sample - - 3 µL




CALCULATION

LDL - C Concentration (mg/dL) =

(OD2-OD1) Sample

--------------------------- x Calibrator Concentration

(OD2-OD1) Calibrator

ALTERNATIVE MEASUREMENT



Wavelength I 546 nm


Temperature 370C


Linearity 450 mg/dL

Blank Reagent

Incubation time 5 min +5 min

Sample volume 5 µL







Sample - - 5 µL

Mix and incubate for 5 minutes at 370C.


Mix and incubate for 5 minutes at 370C. Measure the absorbance of calibrator &sample against reagent blank at 546 nm/630 nm.

CALCULATION


LDL - C Concentration (mg/dL) = ------------------------------- x Calibrator Concentration

Absorbance of Calibrator





Haemoglobin up to 500 mg/dL


*(when triglyceride in a sample exceeds 1000 mg/dL, dilute the sample 1+9 with saline,repeat the assay and multiply result by 10)

BIBLIOGRAPHY

1. Crouse, J.R., et al.; Studies of Low Density Lipoprotein molecular weight in humanbeing with coronary artery disease. J.Lipid Res 26:5666 (1985)

LDL CHOLESTEROL (D) WITH CALIBRATOR2 x 40 mL, 2 x 60 mL

11415003, 11415004


25

INTENDED USE

This reagent is intended for in vitro quantitative determination of lipase in humanserum or plasma.

- Methyl resorufin method


- Reagent is ready for use


Lipase is a pancreatic enzyme necessary for the absorption and digestion ofnutrients that catalyses the hydrolysis of glycerol esters of fatty acids. Determinationof lipase is used for diagnosis of diseases such as acute and chronic pancreatitisand obstruction of the pancreatic duct. Clinical diagnosis should not be made on asingle test result; it should integrate clinical and other laboratory data.

PRINCIPLE

In the presence of colipase and bile acids lipase splits the synthetic substrate (1,2-O-Dilauryl-rac-glycero-3-glutaricacid (6-methyl-resorufin-ester) to glycerol andmethylresorufin-ester, which is spontaneously degraded to glutaric acid andmethylresorufin. The rate of methylresorufin formation, measured photometricallyis proportional to the catalytic concentration of lipase present in the sample.

REAGENT COMPOSITION

LIPASE (S.L) R1 2 x 10 mL

Goods Buffer (pH 8.0) 40 mmol/L

Taurodeoxycholate 3.4 mmol/L

Deoxycholate 6.4 mmol/L


Colipase 1.7 mg/dL

LIPASE(S.L) R2 1 x 5 mL

Tartrate Buffer (pH 4.0) 1.5 mmol/L

Taurodeoxycholate 3.4 mmol/L

Color substrate 0.13 mmol/L

LIPASE CALIBRATOR 1 x 3 mL

Lipase calibrator concentration is stated on the vial label.

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C, protected from light.

LINEARITY


If the concentration is greater than linearity (300 U/L), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum / Plasma : Up to 60 U/L


Lipase R1 and Lipase R2 are ready to use.

Calibrator : Reconstitute with 3 mL of distilled water. Dissolve the content of thevial by swirling gently to avoid the formation of foam.

Stability : Reconstituted calibrator is stable only for 7 days at 2-80C and 3 monthsat -200C.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean dry disposablepipette tips for dispensing. Close reagent and calibrator bottles immediately afterUse. Avoid direct exposure of reagent to light.

SAMPLE

Serum or plasma with sodium citrate, EDTA or heparin.

LIPASE(S.L)1 x 25 mL

11417001


Mode of Reaction Kinetic Fixed Time




Calibrator concentration As on the vial As on the vial


Blank Reagent Reagent

Delay time 120 sec. 120 sec

No of reading 2 -



Reagent volume 1250 µL (1000+250) 1250 µL (1000 + 250)

Cuvette 1cm light path 1 cm light path



Reagent 1 1000 µL 1000 µL 1000 µL


Sample - - 20 µL

Dist. water 20 µL - -

Mix carefully (do not vortex); incubate for 1-5 minutes at 370C. Then add


Mix and incubate for 2 min at 37oC, read absorbance against reagent blank. Measurethe change in absorbance per minute ( OD/min) during 2 min.

or

Mix and read the optical density (T1) 120 seconds after the Reagent 2 addition. Take

second reading (T2) exactly after 120 seconds.

CALCULATION

Lipase U/L =

( OD/min) Sample - ( OD/min) Blank

---------------------------------------------------- x Calibrator concentration

( OD/min) calibrator - ( OD/min) Blank

or

(T2-T

1 ) of sample

------------------------- x Calibrator concentration

(T2-T

1 ) of standard

BIBLIOGRAPHY

1. Mc Neely , M. ; Lipase. Kaplan, A. et al.; Clin. Chem. The C.V.Mosby Co. St Louis,Toronto. Princeton 1984, 1130-1135

2. Burtis, A., et al. ; Tietz Textbook of Clinical chemistry, 3rd ed AACC3. Neumann, U., et al.; Methods of Enzymatic Analysis, Vol 4, 3rd Ed.


26

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGOT in serum orplasma.

- IFCC recommended procedure




It is present in most of the tissues. Especially in cardiac muscle, liver cells, skeletalmuscle & kidneys. Injury to these tissues results in the release of the enzyme in bloodstream.

Increased levels are found in myocardial infarction. The duration & extent of increaseis related to the infract. GOT determination is of considerable value to differentiatemyocardial infarction from other cardiac disorders.

Increased levels are also found in various types of liver disease, skeletal muscle trauma& in renal diseases.


PRINCIPLE


AST

L- Asparate + alpha - ketoglutarate ----- > Oxaloacetate + L-Glutamate.

MDH

Oxaloacetate + NADH + H+ -------- > L- Malate + NAD+

AST: Aspartate aminotransferase.


REAGENT COMPOSITION

SGOT (S.L) R1 2 x 24 mL / 3 x 40 mL / 4 x 100 mL



LDH > 1500 U/L

MDH > 900 U/L

SGOT (S.L)R2 2 x 6 mL / 3 x 10 mL / 4 x 25 mL

alpha -ketoglutarate 12 mmol/L

NADH 0.24 mmol/L



LINEARITY


If the concentration is greater than 350 U/L, follow the high linearity procedure toget higher linearity of 1000 U/L.

If the concentration is greater than linearity, dilute the sample with normal saline andrepeat the assay. Multiply the result with dilution factor.

NORMAL RANGE





Mix 4 volumes of Reagent 1 (R1 ) with 1 volume of Reagent 2 (R2)


NOTE: Discard the working reagent if the blank absorbance is less than 1.0 at 340 nm.

PRECAUTION



SAMPLE



Normal procedure High Linearity procedure


Slope of reaction Decreasing Decreasing



Factor 1745 1745



Delay 60 sec 60 sec

No of reading 3 3







Sample 100 µL


High Linearity Procedure

Mix and incubate at for 1 minutes 370C. Read the change in absorbance per 20 sec,during 1 minute.

CALCULATION

SGOT activity (U/L) = ( OD/min) x 1745

BIBLIOGRAPHY

1. Clin. Chem, Acta. 70, 19-42 (1976)2. Thefeld, W., et al.; Dtsch. Med Wschr.99, 343 (1974)

SGOT (S.L) 2 x 30 mL, 3 x 50 mL, 4 x 125 mL

11408005, 11408003, 11408007

27

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGPT in serum orplasma.

- IFCC recommended methodology




It is present in most of the tissues, but mainly found in the liver. Increased levels arefound in hepatitis, cirrhosis, obstructive jaundice & other hepatic disease. SGPT activityis markedly elevated even before clinical signs of jaundice become apparent in diseaseassociated with hepatic necrosis. Slight elevations are also found in myocardialinfarction.

PRINCIPLE

Kinetic determination of Alanine Aminotransferase (ALT) according to the followingreaction.

ALT

L-Alanine + alpha-ketogutarate ----- >Pyruvate +L-Glutamate

LDH

Pyruvate +NADH+ H+ ----- > L-Lactate +NAD+



REAGENT COMPOSITION

SGPT (S.L) R1 2 x 24 mL / 3 x 40 mL / 4 x 100 mL



LDH > 1500U/L

SGPT (S.L)R2 2 x 6 ml / 3 x 10 mL / 4 x 25 mL

alpha-ketoglutarate 16 mmol/L

NADH 0.24 mmol/L



LINEARITY


If the concentration is greater than 350 U/L, follow the high linearity procedure toget higher linearity of 1000 U/L.

If the concentration is greater than linearity dilute the sample with normal saline andrepeat the assay. Multiply the result with dilution factor.

NORMAL RANGE





Mix 4 volumes of Reagent 1 (R1) with 1 volume of Reagent 2 (R2)



PRECAUTION



SAMPLE



Normal procedure High Linearity procedure


Slope of reaction Decreasing Decreasing



Factor 1745 1745



Delay 60 sec 60 sec

No of reading 3 3







Sample 100 µL


High Linearity Procedure

Mix and incubate for 1 minute at 370C. Read the change in absorbance per 20 secduring 1 minutes.

CALCULATION

SGPT activity (U/L) = ( OD/min) x 1745

BIBLIOGRAPHY

1. Clin. Chem, Acta. 105, 147-172 (1780)2. Thefeld, W., et al.; Dtsch. Med Wschr.99, 343 (1994)

SGPT (S.L) 2 x 30 mL, 3 x 50 mL, 4 x 125 mL

11409005, 11409003, 11409006

28

INTENDED USE

This reagent is intended for in vitro quantitative determination of triglycerides in serumor plasma.

- GPO-PAP methodology


- Contains LCF (Lipaemic clearing factor) which minimizes rerun


Triglycerides are simple lipids, formed in the liver by glycerol & fatty acids. Theyare transported by VLDL, LDL & constitute about 95% of fat, stored as source ofenergy in the tissue & plasma.

Increased levels are found in hyperlipidemias, diabetes, nephrotic syndrome &hypothyroidism. Increased levels are risk factor for arteriosclerotic coronary disease,peripheral vascular disease, acute pancreatitis & hyperlipoproteinaemia. Decreasedlevels are found in malnutrition & hyperthyroidism.

PRINCIPLE

Enzymatic determination of triglyceride is based on following reactions:

LPL

TGL+H2O ------ > Glycerol + Fatty acid

GK

Glycerol + ATP ------ > Glycerol-3-phosphate + ADP

Mg++

GPO

Glycerol-3-phospahte+O2

------- > Dihydroxyacetone phosphate +H2O

2

POD

2H2O

2+4-Aminoantipyrine+ p-chlorophenol ------- > Red Quinoneinimine

GPO = Glycereol-3-phosphate Oxidase



REAGENT COMPOSITION

TRIGLYCERIDES REAGENT 4x10mL / 5x25mL / 6x50mL / 5x100mL



Potassium ferrocynate 10 mmol/L

Magnesium Salt 17 mmol/L

4-Aminoantipyrine 0.9 mmol/L

ATP 3.15mmol/L

Lipoprotein Lipase > 1800 U/L


Glycerol – 3- phosphate oxidase > 3500 U/L


TRIGLYCERIDES STANDARD 1 x 4 mL

Triglycerides std.concentration 200 mg/dL



LINEARITY



REFERENCE RANGE



Male : 60-165 mg/dL

Female : 40-140 mg/dL


The reagent ready to use.

PRECAUTION



SAMPLE





Wavelength I 505 nm (492-550 nm)


Temperature 370C


Linearity 1000 mg/dL

Blank Reagent








Standard - 10 µL -

Sample - - 10 µL


CALCULATION


Triglycerides Con. (mg/dL) = ------------------------------ x 200


BIBLIOGRAPHY

1. Schettler, G., Nussel, E.; Arav. Med 10, 25 (1975)2. Jacobs, N.J. , VanDemark, P.J.; Arch, Biochem, Biophy. 88, 250 – 255 (1960)

TRIGLYCERIDES (S.L)4x 10 mL, 5 x 25 mL, 6x 50 mL, 5 x 100 mL

11410007, 11410002, 11410008, 11410004


29

INTENDED USE

This reagent is intended for in vitro quantitative determination of urea in serum, plasma& urine.

- Urease / GLDH methodology


- Working reagent can be prepared as per requirement


Proteins cannot be stored in human body, so excess should be broken down. Aminoacids which from the components of proteins, break down to give ammonia. This istoxic & so through a series of chemical reactions (urea cycle) non toxic urea isproduced & this is released into the blood which is filtered in the kidney & excretedin the urine.


Decreased levels are found in liver failure & pregnancy.

PRINCIPLE

Enzymatic determination of Urea according to the following reaction.

Urease

Urea + H2O ------------> 2NH

3 + CO

2

GLDH

2 NH3

+2- ketoglutarate + 2NADH ----------> L-Glutamate+2NAD+ + 2H2O

REAGENT COMPOSITION

UREA U.V (S.L) R1 2 x 24 mL/ 2 x 40 mL/ 2 x 100 mL


ADP 0.7 mmol/L


Urease > 6500 U/L

GLDH > 1100 U/L

UREA U.V (S.L) R2 2 x 6 mL/ 2 x 10 mL / 2 x 25 mL

NADH 0.25 mmol/L

2-Oxoglutarate 5 mmol/L

UREA U.V STANDARD 1 x 4 mL

Standard concentration for Urea 50 mg/dL

STORAGE & STABILITY


LINEARITY



REFERENCE RANGE



Serum/ Plasma : 10-50 mg/dL

Urine : 20-35 gm/24 hr


Mix 4 volume of Reagent 1 (R1) with 1 volume of Reagent 2 (R2)

Working reagent is stable for 30 days at 2-80c.

Note

Discard the working reagent if the blank absorbance is less than 1.0 at 340 nm.

PRECAUTION

To avoid contamination useclean laboratory wares. Avoid direct exposure of reagentto light.

SAMPLE

Serum, Plasma ( free of hemolyses). Do not use anticoagulants containing Fluorideor Ammonium Ions.

Urine(1/100 diluted with distilled water (DI water). Multiply result with dilution factor.

UREA U.V (S.L)2 x 30 mL, 2 x 50mL, 2 x 125 mL

11412007, 11412002, 11412008


Mode of Reaction Fixed Time


Wavelength 340

Temperature 370C

Blank DI Water


Linearity 300 mg/dL

Delay time 30 sec

Interval 60 sec





Standard Sample

Working Reagent 1000 µL 1000 µL

Standard 10 µL -

Sample - 10 µL


addition. Take second reading (T2) exactly 60 seconds after the first reading.

CALCULATION

( T1 –T

2)of Sample

Urea Conc. (mg/dL) = ------------------------ x 50

(T1-T

2) of standard

(T1 –T

2) of Sample

Urea BUN Conc. (mg/dL) = ------------------------ x 23.4

(T1-T

2) of standard

BIBLIOGRAPHY

1. Kassirer, J.P. New eng. J. Med.285, 385 (1971)2. Talke, H., Schubert, G.E. ; Klin. Wochenschr, 43, 174(1965)


30

INTENDED USE:

This reagent is intended for in vitro quantitative determination of uric acid in serum,plasma & urine.

- Uricase – PAP methodology


- Fast incubation, just 5 minutes at 370C


Uric acid is the end product of purine metabolism. Uric acid is excreted by the kidneys.Increased levels are found in Gout, arthritis, impaired renal functions & starvation.Decreased levels are found in yellow atrophy of the liver.

PRINCIPLE

Enzymatic determination of uric acid according to the following reactions.

Uricase

Uric acid + 2H2O+O

2---------- > Allantoine +CO

2 +H

2O

2

Peroxidase

2H2O

2 + 4 -Aminoantipyrine +EHSPT ---------------- > Red quinone

EHSPT = N-Ethyl N-(2-Hydroxy-3-Sulfoproyl) n-Toluidine

REAGENT COMPOSITION

URIC ACID (S.L) R1 2 x 30 mL / 2 x 50 mL / 2 x 100 mL


EHSPT 1.10 mmol/L

Ferrocyanure 50 µmol/L

Amino-4-antipyrine 0.37 mmol/L

Uricase > 140 U/L


URIC ACID STANDARD 1 x 4 mL

Uric acid standard concentration 8 mg/dL



LINEARITY



NORMAL RANGE



Serum / Plasma

Men : 3.4 - 7.0 mg/dL

Women : 2.4 - 5.7 mg/dL


The reagent is ready to use

Note : Discard the reagent if the blank absorbance exceeds 0.3

PRECAUTION



SAMPLE

Serum / EDTA or Heparin plasma (free of haemolysis)

URIC ACID (S.L)2x30mL, 2x50mL, 2x100mL

11413004, 11413002, 11413003




Wavelength I 546 nm (540 - 560)


Temperature 370C


Blank Reagent

Incubation time 5 minutes

Linearity 25 mg/dL






Reagent 1000 µL 1000 µL 1000 µL

Standard - 25 µL -

Sample - - 25 µL

Mix and incubate 5 min. at 370C. Measure absorbance of sample and standard againstthe reagent blank.

CALCULATION


Uric Acid Con. (mg/dL) = ------------------------------- x 8


BIBLIOGRAPHY

Barham, D., Trinder; Analyst 97, 142(1972)


31

INTENDED USE

This reagent is intended for in vitro quantitative determination of albumin in serumor plasma.

- Bromocresol green methodology

- Linear up to 6 g/dL


Albumin which is synthesized in the liver constitutes a major part of the totalproteins in the body, the other part being globulin, they form the major portion ofthe dissolved substances in the plasma. Functions of albumin includes distributionof extracellular fluid, regulation of osmotic pressure, acts as transport agent for awide variety of substance such as hormones, lipids, vitamins etc.

Increased levels are seen in dehydration.

Decreased levels are seen in liver disease (Hepatitis, Cirrhosis), malnutrition, kidneydisorders, increased fluid loss during extensive burns & malabsorption.

PRINCIPLE

The reaction between albumin from serum or plasma and the dye bromocresol-green produces a change in colour that is proportional to the albumin concentration.

REAGENT COMPOSITION

ALBUMIN REAGENT 4 x 50 mL

Succinate Buffer (pH 4.20) 75 mmol/L

Bromocresol green 0.14 g/L

ALBUMIN STANDARD 1 x 3 mL

Albumin standard concentration 3 g/dL

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat room temperature & standard at 2 - 80C.

LINEARITY

This reagent is linear up to 6 g/dL.

If the concentration is greater than linearity (6 g/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum / plasma 3.5 – 5.5 g/dL


Reagent and standard are ready to use.

PRECAUTION



SAMPLE





Wavelength 630 nm

Temperature 30oC

Standard Concentration 3 g/dL

Linearity 6 g/dL

Blank Reagent



Reagent volume 1000µL





Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 1 minute. Measure absorbance of standard & sample againstthe reagent blank.

ALBUMIN4 x 50 mL

11001003

CALCULATION

Absorbance of SampleAlbumin Con. (g/dL) = -------------------------------- x 3


BIBLIOGRAPHY

1. Doumasa, B.T., et al; Clin. Chim Acta 31, 87 pp (1971)

2. Weis, W. A. ; Klin. Wochenschr. 43, S.273 (1965)


32


Sample Blank Test

Direct bilirubin reagent 1000 µL 1000 µL

Activator Direct - 20 µL

Serum 50 µL 50 µL

Mix well and incubate for 5 minutes at room temperature. Measure the absorbanceof test against respective Blank at 546 nm.

CALCULATION

With factor :

Direct Bilirubin = OD of test – OD of sample blank x16

With calibrator :

OD of test –OD of sample blank

Bilirubin Concentration =---------------------------------------- x concentration of std.OD of calibrator – OD of calibrator blank

Alternative Method – without sample blank




Wavelength I 546 nm


Temperature 30oC

Factor (Direct ) 20.0

Blank Reagent

Linearity 20 mg/dL

Reaction time 5 min



Activator 20 µL



Reagent Blank Test

Direct Bilirubin Reagent 1000 µL 1000 µL

Activator Direct 20 µL 20 µL

Serum / Calibrator - 50 µL

Mix well and incubate for exactly 5 minutes. Measure the absorbance of calibratorand test against reagent blank at 546/630 nm.

CALCULATION

With factor :

Direct Bilirubin = OD of test – OD of reagent blank x 20

With calibrator :

OD of test –OD of reagent of blank

Bilirubin Concentration =------------------------------------------- x concentration of std.OD of calibrator – OD of sample blank

BIBLIOGRAPHY

1. Water, M., Gerard, H.; MICROCHEM JM 15, 231(1980)2. Annino J. S.; C.C. Principles and procedure,19603. A.A. A.C.C.; Clin. Chem. 8 : 405,196

INTENDED USE

This reagent is intended for in vitro quantitative determination of Bilirubin in serumor plasma .


- Fast incubation 5 minutes at room temperature

- Sample volume only 50 µL

- Without sample blank procedure also included


Bilirubin is formed by the break down of RBC’s in the spleen, liver & bone marrow.Small amount of bilirubin circulates in the plasma loosely bound to albumin, whichis not water soluble. This is referred to as indirect or unconjugated bilirubin. In theliver bilirubin is conjugated with glucuronic acid, which forms a soluble compound.This is referred to a direct bilirubin.

Elevated levels are found in Hepatitis, Cirrhosis, Haemolytic jaundice, obstruction ofbiliary tract & drug induced reactions.

PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. DirectBilirubin reacts with diazotized sulfanilic acid to form azobilirubin.

REAGENT COMPOSITION

DIRECT BILIRUBIN REAGENT 4 x 50 mL

Sulfanilic acid 28.9 mmol/L

Hydrochloric acid 165 mmol/L

Preservatives and stabilizers

DIRECT BILIRUBIN ACTIVATOR 2 x 4 mL

BILIRUBIN CALIBRATOR

Not provided along with the Kit, recommended Agappe multicalibrator product code: 11610001


The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. The standard & activator should be stored at 2 - 80C

LINEARITY



NORMAL RANGE



Direct Bilirubin up to 0.4 mg/dL


Reagents are ready to use.

PRECAUTION



SAMPLE





Wavelength 546 nm

Temperature 30oC

Factor (Direct ) 16.0

Blank Sample blank

Linearity 20 mg/dL

Reaction time 5 min



Activator 20 µL


BILIRUBIN DIRECT4 x 50 mL

11003003


33

INTENDED USE

This reagent is intended for in vitro quantitative determination of Bilirubin in serumor plasma.

- Modified TAB method



- Sample volume only 50 µL

- Without sample blank procedure also included


Bilirubin is formed by the break down of RBC’s in the spleen, liver & bone marrow.Small amount of bilirubin circulates in the plasma loosely bound to albumin, whichis not water soluble. This is referred to as indirect or unconjugated bilirubin. In theliver bilirubin is conjugated with glucuronic acid, which forms a soluble compound.This is referred as direct bilirubin.

Elevated levels are found in Hepatitis, Cirrhosis, Haemolytic jaundice, obstruction ofbiliary tract & drug induced reactions.

PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. TotalBilirubin reacts with diazotized sulfanilic acid in the presence of TAB form azobilirubin.

REAGENT COMPOSITION

TOTAL BILIRUBIN REAGENT 4 x 50 mL


TAB 9 mmol/L

Preservatives and Stabilizers

TOTAL BILIRUBIN ACTIVATOR 2 x 4 mL

BILIRUBIN CALIBRATOR

Not provided along with the Kit, recommended Agappe multicalibrator product code: 11610001


The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. The Calibrator & activator should be stored at 2 - 80C.

LINEARITY



NORMAL RANGE



Total Bilirubin - up to 1.2 mg/dL



PRECAUTION



SAMPLE

Serum/Plasma (free of haemolysis)




Wavelength 546 nm

Temperature 30oC

Factor (Total ) 25

Blank Sample blank

Linearity 25 mg/dL

Reaction time 5 min



Activator 20 µL


BILIRUBIN TOTAL-TAB4 x 50 mL

11003005


Sample Blank Test

Total Bilirubin Reagent 1000 µL 1000 µL

Activator Total - 20 µL

Serum / Calibrator 50 µL 50 µL

Mix well and incubate for 5 minutes at room temperature. Measure the absorbanceof calibrator and test against respective Blank at 546 nm.

CALCULATION

With factor :

Total Bilirubin = OD of test – OD of sample blank x 25

With calibrator :

OD of test –OD of sample blank

Bilirubin Concentration = ---------------------------------------------- x Conc.of calib.

OD of calibrator – OD of calibrator blank

Alternative Method – without sample blank




Wavelength I 546 nm


Temperature 30oC

Factor (Total ) 29

Blank Reagent blank

Linearity 25 mg/dL

Reaction time 5 min



Activator 20 µL



Reagent Blank Test

Total bilirubin reagent 1000 µL 1000 µL

Activator Total 20 µL 20 µL

Serum / Calibrator - 50 µL

Mix well and incubate for exactly 5 minutes.Measure the absorbance of calibrator andtest against reagent blank at 546/630 nm.

CALCULATION

With factor :

Total Bilirubin = OD of test – OD of reagent blank x 29.00

With calibrator :

OD of test – OD of reagent of blank

Bilirubin concentration = ----------------------------------------------- x conc. of calib.

OD of calibrator – OD of sample blank

BIBLIOGRAPHY

1. Walter, M., Gerard, H.; MICROCHEM JM 15, 231.(1980)2. Annino J. S.; C.C. Principles and procedure,19603. A.A. A.C.C.; Clin. Chem. 8 : 405,196

ADL/V.02/JAN 2013

34

INTENDED USE


- Modified DMSO method



- Sample volume only 50µL



Elevated levels are found in Hepatitis, Cirrhosis, Haemolytic jaundice, obstructionof biliary tract & drug induced reactions.

PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. TotalBilirubin reacts with diazotized sulfanilic acid in the presence of DMSO to formazobilirubin.

REAGENT COMPOSITION



HCl 165 mmol/L

DMSO 7 mmol/L



BILIRUBIN ARTIFICIAL STANDARD 1 x 4 mL

Total bilirubin standard concentration10mg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. The standard & activator should be stored at 2 - 80C.

LINEARITY



REFERENCE RANGE



Cord : 2.0 mg/dL

0-1 day : 1.4-8.7 mg/dL

1-2 days : 3.4-11.5 mg/dL

3-5 days : 1.5-12.0 mg/dL

Adult : 0.3-1.2 mg/dL



PRECAUTION



SAMPLE


BILIRUBIN TOTAL4 x 50 mL

11003002




Wavelength 546 nm/ 532 nm

Temperature 30oC

Factor (Total ) 20.5(546nm)/23.5 (532 nm)

Blank Sample blank

Linearity 20 mg/dL

Reaction time 5 min



Activator 20 µL



Sample Blank Test

Total Bilirubin Reagent 1000 µL 1000 µL

Activator Total - 20 µL

Serum / Calibrator 50 µL 50 µL

Mix well and incubate for 5 minutes at room temperature. Measure the absorbanceof calibrator and test against respective Blank at 546 nm.

CALCULATION

With factor :

Total Bilirubin = OD of test – OD of sample blank x Factor

With artificial standard :

OD of test –OD of sample Blank

Bilirubin Concentration =--------------------------------------- x 10

OD of standard

BIBLIOGRAPHY

1. Walter, M., Gerard, H.; MICROCHEM JM 15, 231.(1980)2. Annino J. S.; C.C. Principles and procedure,19603. A.A. A.C.C.: Clin. Chem. 8 : 405,19


35

INTENDED USE


- Modified DMSO/DIAZO method- Linear up to 20 mg/dL- Fast incubation 5 minutes at room temperature- Sample volume only 50 µL




PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. TotalBilirubin reacts with diazotized sulfanilic acid in the presence of DMSO to formazobilirubin.

REAGENT COMPOSITION

TOTAL BILIRUBIN REAGENT 2 x 50mL

Sulfanilic acid 28.9mmol/L

HCl 165mmol/L

DMSO 7mmol/L





HCl 165 mmol/L


BILIRUBIN ARTIFICIAL STANDARD 1 x 4 mL

Total bilirubin standard concentration 10 mg/dL

Direct bilirubin standard concentration 7.7 mg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. The standard & activator should be stored at 2 - 80C.

LINEARITY



NORMAL RANGE




Direct bilirubin up to 0.4 mg/dL



PRECAUTION



SAMPLE


BILIRUBIN TOTAL & DIRECT4 x 50 mL

11003001





Temperature 30oC

Factor (Total ) 20.5(546nm)/23.5 (532 nm)

Factor (Direct) 16(546nm)/18 (532 nm)

Blank Sample blank

Linearity 20 mg/dL

Reaction time 5 min



Activator 20 µL



Total Bilirubin Direct Bilirubin

Sample blank Test Sample blank Test

T.Bilirubin reagent 1000 µL 1000µL - -

D.Bilirubin reagent - - 1000 µL 1000 µL

Activator (T/D) - 20 µL - 20 µL

Serum 50 µL 50 µL 50 µL 50 µL

Mix well and incubate for exactly 5 minutes. Measure the absorbance of the sampleagainst the respective sample blank at 546 or 532 nm.

CALCULATION

For semi auto with factor :

Total Bilirubin = OD of test – OD of sample blank x Factor

Direct Bilirubin = OD of test – OD of sample blank x Factor

With artificial standard :


Total Bilirubin Concentration = ------------------------------------------ x 10OD of standard


Direct Bilirubin Concentration = ------------------------------------------ x 7.7 OD of standard

BIBLIOGRAPHY

1. Walter, M., Gerard, H.; MICROCHEM J M 15, 231.(1980)2. Annino J. S.; C.C. Principles and procedure, 1960.3. A.A. A.C.C.; Clin. Chem. 8 : 405,19

ADL/V.02/JAN 2013

36

INTENDED USE

This reagent is intended for in vitro quantitative determination of calcium in serum,plasma & urine.

- Modified OCPC methodology



Calcium is an important ion present in the body. Mainly it is found in bones. In serumcalcium exists equally in a free ionized form & also in a bound form with albumin.Calcium helps in enzyme activation, muscle contraction, coagulation of blood,regulation of some hormonal secretions & cell membrane permeability.

Increased levels are found in hyperthyroidism, malignant tumors, acute osteoporosis& adrenal insufficiency.

Decreased levels found in hypoparathyrodism, osteomalacia, rickets, renal failure &tetanus.

PRINCIPLE

Calcium OCPC procedure is based on on the reaction of calcium ions (Ca 2++) with O-cresolphthalein complex in an alkaline solution to form an intense violet colouredcomplex which shows maximum absorbance at 578nm. the 8-hydroxy quinoloineprevents Mg2++ interference upto 4 mmol/L.

REAGENT COMPOSITION

CALCIUM DYE REAGENT (R2) 2 x 25 mL / 2 x 50 mL

Diethylamine 360 mmol/L

CALCIUM BASE REAGENT (R1) 2 x 25 mL / 2 x 50 mL

O-Cresolphthalein complex 0.15 mmol/L

8-Hydroxyquinoline 17.2 mmol/L

CALCIUM STANDARD 1 x 4 mL

Calcium standard concentration 10 mg/dL

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum : 8.8 - 10.2 mg/dL

Urine : 100 - 400 mg/24 hrs


Mix reagent 1 (R1) and Reagent 2 (R2) in the ratio 1:1.

PRECAUTION



(Use acid washed (50 % HNO3) glass wares & tips)

SAMPLE

Serum / plasma (free of haemolysis) / Urine (1/3 diluted)




Wavelength 578 nm (565-580 nm)

Temperature 30oC


Linearity 15 mg/dL

Blank Reagent





CALCIUM4 x 25 mL, 4 x 50 mL

11005001, 11005002




Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 5 min. at room temperature. Read the absorbance of standardand sample against reagent blank.

CALCULATION


Calcium Conc. (mg/dL) = --------------------------------- x 10


INTERFERENCE

Bilirubin concentrations higher than 20 mg/dL and phosphate higher than 40 mg/dL,will interfere with the assay.

BIBLIOGRAPHY

1. Schwarzenbach, G.; Analyst., 80, (1955) 713-7292. Kessler, G., Wolfman, M., Clin.Chem., 10, (1964) 686 – 7033. Connerty, H. V., Briggs, A.R., Am. J. Clin.Pathol., 45, (1965) 290-2964. Gitelmann, H. J. Anal Biochem 18, (1967) 521-5315. Biggs, H.G., Moorehead, W. R.; Clin.Chem., 20, (1974) 1458-1460


37

INTENDED USE

This reagent is intended for in vitro quantitative determination of calcium in serum,plasma & urine.

- Modified Arsenazo III method

- Linear up to 18mg/dL



Increased levels are found in hyperthyroidism, malignant tumors, acute & osteoporosisadrenal insufficiency.

Decreased levels found in hypoparathyrodism, osteomalacia, rickets, renal failure &tetanus.

PRINCIPLE

At a neutral pH the Ca2+ form with Arsenazo III a complex, the color intensity of whichis directly proportional to the concentration of calcium in the sample.

REAGENT COMPOSITION

CALCIUM ARSENAZO REAGENT 4 x 25 mL

MES, pH 6.50 100 mmol/L

Arsenazo III 200 mmol/L

CALCIUM ARSENAZO STANDARD 1 x 4 mL

Calcium Standard Concentration 10 mg/dL

STORAGE & STABILITY


LINEARITY


If the concentration is greater than linearity (18mg/d/L), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum / plasma : 8.8 - 10.2 mg/dL



PRECAUTION



NOTE: Use acid washed (50% HNO3) glass wares & tips.

SAMPLE


Take dilution factor into account for the calculation of the concentration in urine.




Wavelength 630 or 650 nm

Temperature RT


Linearity 18 mg/dL

Blank reagent





CALCIUM (ARSENAZO)4 x 25 mL

11006001




Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 1 minute. Measure the absorbance of standard and sampleagainst reagent blank.

CALCULATION


Calcium Conc. (mg/dL) = ------------------------------- x 10


BIBLIOGRAPHY

Baver, P. J.; Anal. Biochem., 110, (1981), 61

Biggs, H.G., Moorchand, W.R., (1974), cLIN. CHEM. 20, 1458-1460.


38

INTENDED USE

This reagent is intended for in vitro quantitative determination of Chloride in serum,plasma & urine.

- Modified Thiocyanate method

- Linear up to 130 mEq/L


Chloride & bicarbonate are the principle anions (-vely charged) whereas sodium &potassium are the principle cations (+vely charged) in the plasma. Chloride ions areinvolved in regulation of water distribution between the tissues by maintainingosmotic pressure & normal cation and anion balance between intra & extra cellularfluids.

Elevated levels are seen in conditions like dehydration & congestive cardiac failure.

Decreased levels are seen in condition such as salt losing nephritis, diabetic acidosis& renal failure.

PRINCIPLE

In an acid medium chloride ions and mercury – II – thiocynate form thiocynate ions.These ions react with HNO3 and iron-III-ions and effect a red color. The intensity ofthe color is directly proportional to the concentration of chloride ions.

REAGENT COMPOSITION

CHLORIDE REAGENT 4 x 50 mL

Mercuric (II) thiocyanate 2 mmol/L

Nitric acid 29 mmol/L

Ferric Nitrate 20 mmol/L

CHLORIDE STANDARD 1 x 4 mL

Chloride standard concentration 100 mEq/L

STORAGE & STABILITY


LINEARITY

This reagent is linear up to (130 mEq/L).

If the concentration is greater than linearity (130 mEq/L), dilute the sample withDistilled water and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE


The following value may be used as guideline.

Serum : 97 - 108 mEq/L

Urine : 120 - 240 mEq/L/24 hr



PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / Urine (Dilute sample 1:1 with distilled water andmultiply the result with 2)




Wavelength 505 nm (480-550nm)

Temperature RT

Standard Concentration 100 mEq/L

Linearity 130 mEq/L

Blank Reagent

Reaction time 1 min




CHLORIDE4 x 50 mL

11007001




Standard - 10 µL -

Sample - - 10 µL

Mix and incubate for 1 min. Measure the absorbance of standard and sample againstreagent blank.

CALCULATION


Chloride conc. (mEq/L) = ------------------------------ x 100


BIBLIOGRAPHY

Schonfeld, R. G., Lowellen, C. S.; Clin. Chem. 10, 533 pp (1964)


39

INTENDED USE

This reagent is intended for in vitro quantitative determination of Copper in serum.


Copper is an essential trace mineral in humans, the function of copper is to help torelease energy, helps in melanin production in the skin, helps in the production ofred blood cells and aid in the absorption and transport of iron.

Acquired copper deficiency can cause hematological/neurological manifestations.Wilson disease (copper toxicity) is associated with neurological manifestations andlow serum copper, with copper deposited in tissues responsible for the toxicity. Thesymptoms of acute copper poisoning include nausea, vomiting and abdominal andmuscle pain.

PRINCIPLE

Copper in an acid medium reacts with the chromogen Di-Br-PAESA 4-(3,5-dibromo-2- pyridylazo)-N-ethyl-N-(3-sulphopropyl0aniline.) to form a coloured complex.Intensity of the colour is directly proportional to the amount of Copper present inthe sample.

REAGENT COMPOSITION

COPPER REAGENT 1 1 x 25 mL

Acetate buffer(pH 4.9) 0.1 M

COPPER REAGENT 2 1 x 25 mL

3,5 Di-Br-PAESA

Preservatives

COPPER STANDARD 1 x 4 mL

Copper Standard (Concentration) 200 µg/dL



Note:Reagent 1 solidifies when kept at 2-8oC. Allow the reagent to melt by keeping atroom temperature before use.

LINEARITY

This reagent is linear up to 500 µg/dL.

If the concentration is greater than linearity (500 µg/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

REFERENCE VALUES



Men : 70 - 140 µg/dL

Women : 80- 155 µg/dL

Newborns : 20- 70 µg/dL

Children up to 6 years : 90-190 µg/dL

Children up to 12 years : 80-160 µg/dL

PREPARATION OF WORKING REAGENT

Mix in equal parts the reagent 1 & reagent 2 .

PRECAUTION

To avoid contamination, use clean laboratory wares. It is recommended to usedisposable tubes. Use clean, dry disposable pipette tips for dispensing. Close reagentand standard bottles immediately after use. Avoid direct exposure of reagent to light.

Note: Use acid washed (50% HNO3) glass wares and tips.

SAMPLE

Serum or Plasma (free haemolysis). Use heparin as anticoagulant.Highly lipemic serummay interfere the assay. it is recommended to filter or centrifuge the sample




Wavelength 580 nm

Temperature 30oC

Standard concentration 200 µg/dL

Linearity 500 µg/dL

Blank Reagent Blank

Incubation Time 10 min






WorkingReagent 1000 µL 1000 µL 1000 µL

Distilled water 70 µL - -

Standard - 70 µL -

Sample - - 70 µL

Mix and incubate for 10 minutes at 30oC. Read the absorbance(A) of standard andsample against blank at 580 nm. The colour is stable for 30 minutes.

CALCULATION


Copper µg/dL = -------------------------------- x 200


BIBLIOGRAPHY

1. Pasquinelli, F.; Diagnostica e Tecniche di Laboratorio, (pag.:1099-1102) RossiniEditrice.(1984)

2. Akitaabe, Sumico Yiamashita; Clin. Chem. 35(4):197,552-554(1989)3. Cuiti, R, Gallia, Giorn.; It. Chim. Clin. 12(2): 91-100 (1987)

COPPER2 x 25 mL

11020001

40

INTENDED USE

This reagent is intended for in vitro quantitative determination of creatinine in serum,plasma & urine.

- Modified Jaffe’s method


- No Bilirubin interference up to 10 mg/dL


Creatinine is formed in muscles from phospho creatinine. It is an important form ofenergy, being a store of high-energy phosphate. Creatinine determinations have oneadvantage over Urea determination that it is not affected by a high protein diet.

Serum creatinine is more specific & sensitive indicator of renal function. Simultaneousestimations of serum urea & creatinine provides better information. Serum ureanitrogen, creatinine ratio is > 15 in pre renal failure, & < 10 in renal failure.

Decreased levels are found in muscle dystrophy.

PRINCIPLE

Creatinine reacts with picric acid to produce a colored compound, creatinine alkalinepicrate. The change in absorbance is proportional to the creatinine concentration.

REAGENT COMPOSITION

CREATININE BASE REAGENT (R1) 2 x 50 mL

Sodium hydroxide 300 mmol/L

Sodium Phosphate 25 mmol/L

CREATININE DYE REAGENT (R2) 2 x 50 mL

Picric Acid 8.73 mmol/L

Surfactant

CREATININE STANDARD 1 x 4 mL

Creatinine standard concentration 2 mg/dL

STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum : Men : 0.7 – 1.4 mg/dL

Female : 0.6 – 1.2 mg/dL

Urine : 0.80 – 1.80 gm/24 hour


Mix 1 volume of Reagent 1 (R1) with 1 volume of Reagent 2 (R2).

PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / Urine (diluted 1/100 with distilled water)

CREATININE4 x 50 mL

11009001


Mode of Reaction Fixed time



Temperature 370C


Linearity 24 mg/dL

Blank D I water

Delay time 60 sec

Interval 60 sec





Standard Sample

Working Reagent 1000 µL 1000 µL

Standard 100 µL -

Sample - 100 µL


addition. Exactly 60 seconds after the first reading take second reading (T2)

CALCULATION

(T2- T

1) of sample

Creatinine conc. (mg/dL) = ---------------------- x 2

(T2- T

1) of Standard

BIBLIOGRAPHY

1. Allen, L.C.; Clin chem. Vol.28 No.3, 1982, 555.2. Haeckel, R., et al. ; Clin. Chem. 27/1 179-183 (1981).3. Tanganelli, E., Prencipe,L. , Bassi, D., Cambiaghi, S. and Murador, E.; Clin.Chem

28/7, 1461-1464 (1982)

ADL/V.02/JAN 2013

41

INTENDED USE

This reagent is intended for in vitro quantitative determination of Haemoglobin inblood.

- Based on cyanmethaemoglobin method

- Linear up to 20 gm/dL


A decrease in haemoglobin below normal range is an indication of anaemia. An increasein haemoglobin concentration occurs in haemoconcentration due to loss of bodyfluid in severe diarrhea and vomiting. High values are also observed in congenital heartdisease (due to reduced oxygen supply) in emphysema and also in poly cythemia.

Haemoglobin concentration drops during pregnancy due to haemodilution

PRINCIPLE

The Haemoglobin (oxyhaemoglobin,methemoglobin, Carboxyhaemoglobin) isconverted to cyanmethaemoglobin according to the following reactions.

K3 Fe(CN)6

Haemoglobin --------------- > Methemoglobin

KCN

Methemoglobin --------------- > cyanmethemoglobin

The intensity of the color is proportional to haemoglobin concentration and iscompared to known cyan methaemoglobin standard at 540 nm (green filter).

REAGENT COMPOSITION

HAEMOGLOBIN REAGENT 1000 mL

Potassium Phosphate 2.0 mmol/L

Potassium ferricyanide 0.60 mmol/L

Potassium cyanide 0.90 mmol/L

Sodium chloride 1.4 mmol/L

HAEMOGLOBIN STANDARD 1 x 4 mL

Cyanmethaemoglobin standard con. 60 mg/dL



LINEARITY

This reagent is linear up to 20 gm/dL.

REFERENCE RANGE



Cord blood :13.5 - 20.5 gm/dL

0.5 - 2.0 yrs :11.3 - 14.1 gm/dL

Male :13.2 - 17.3 gm/dL

Female :11.7 - 15.3 gm/dL


The reagent is ready to use.‘

PRECAUTION


Avoid direct exposure of reagent to light. Do not pipette the reagent with mouth.

SAMPLE

Fresh whole blood.




Wavelength 546nm (530-550nm)

Temperature 30oC

Blank Reagent

Linearity 20 gm/dL

Standard concentration 15 gm /dL (60x0.251)





*NOTE : Analyzer users directly enter given Factor without running standard.

Factor 35


Blank Sample

Hb Reagent 5000 µL 5000 µL

Sample - 20 µL

Mix well and incubate at room temperature for 5 minutes. Measure the absorbanceof sample against reagent blank and measure the absorbance of standard directlyagainst blank (distilled water).

CALCULATION

Haemoglobin Conc. (gm/dL) =


---------------------------- x 60 x 0.251


Dilution factor

Where, 0.251 = ------------------

Convertion factor

OR


-------------------------------- x 15


15 = 60 x 0.251

BIBLIOGRAPHY

1. Drabkin, D.L., et al.; J.Bio.Chem, 98 (1932), 7192.. Zijlstra, N. C.; Clin.Chem.Acta, 5,(1960) 719

HAEMOGLOBIN 1 x 1000 mL

11011001

ADL/V.02/Jan 2013

3. Tietz Text book of Clin Chem.Carl.AB,Edward.R.A,3rd Edition 1999

42

INTENDED USE

This reagent is intended for in vitro quantitative determination of HDL in serum orplasma.

- Precipitation method, Phosphotungstate magnesium acetate reagent



Lipoproteins are the proteins, which mainly transport lipids in the blood stream. Theyare (HDL) High density lipoproteins, (LDL) Low density lipoproteins, (VLDL) Very lowdensity lipoproteins & chylomicrons. LDL carries cholesterol to the peripheral tissueswhere it can be deposited & increase the risk of atherosclerotic heart & peripheralvascular disease. Hence high levels of LDL are atherogenic. HDL transports cholesterolfrom peripheral tissues to the liver & then for excretion, hence HDL has a protectiveeffect. Hence the determination of serum HDL cholesterol is a useful tool to identifypatients at risk of developing coronary heart disease.

PRINCIPLE

The chylomicrons, Very low density lipoproteins (VLDL) and Low density lipoproteins(LDL) of serum are precipitated by phosphotungstic acid and magnesium ions.

After centrifugation, High density lipoproteins (HDL) are in the supernatant. HDLcontent of supernatant is measured by an enzymatic Method.

REAGENT COMPOSITION

HDL CHOLESTEROL R1 4 x 25 mL

Phosphotungstate 14 mmol/L

Magnesium Chloride 1 mmol/L

Preservative

HDL CHOLESTEROL STANDARD 1 x 4 mL

HDL Cholesterol concentration 50 mg/dL

STORAGE & STABILITY


LINEARITY

The reagent is linear up to 125 mg/dL.


NORMAL RANGE



HDL Cholesterol

Men : 35-55 mg/dL

Women : 45-65 mg/dL

LDL Cholestrol

Suspicious : 150 mg/dL

Elevated : 190 mg/dL


Reagent is ready to use.

PRECAUTION



SAMPLE


HDL CHOLESTEROL4 x 25 mL

11010001




Wavelength I 505 (500 -532 nm)


Temperature 370C


Blank Cholesterol Reagent

Linearity 125 mg/dL






1. PRECIPITATION

Sample 300 µL

HDL reagent 300 µL

Mix well, allow to stand for 10 min. at room temperature, mix again and centrifugefor 10 min, at 4000 rpm.

After centrifugation separate the clear supernatant from the precipitate within 1 hourand determine the HDL Cholesterol concentration using the cholesterol reagent.

2. HDL CHOLESTEROL DETERMINATION :


Cholesterol Reagent 1000 µL 1000 µL 1000 µL

Standard(HDL) - 50 µL -

HDL supernatant - - 50 µL

Mix and incubate for 5 min. at 370C. Measure the absorbance of the standard &sample against the reagent blank.

CALCULATION

HDL Cholesterol Conc. In mg/dL =


------------------------------------ x concentration of standard x 2


where, 2 = dilution factor of the sample.

LDL-Chol conc in mg/dL = Total Cholesterol – (HDL Chol. + Triglycerides / 5)

BIBLIOGRAPHY

1. Assmann, G.; Intermist 20 (1979), 5592. Gordon, T., et al.; Med 62 (1977), 7073. Friedewald, W. T., et al.; Clin.Chem.18 (1972), 499.


43

INTENDED USE

This reagent is intended for in vitro quantitative determination of Phosphorous inserum or plasma.

- Phosphomolybdate methodology



Phosphorous is mainly combined with calcium & is found in bones. It is involved inthe carbohydrate metabolism & is a component of many other substances. Some ofits important functions include maintaining of acid-base balance, skeletal muscleformation. It is also required for normal functioning of RBC’s & muscles.

Increased levels are found in hypothyroidism, renal failure, bone metastasis & liverdisease.

Decreased levels are found in hyperparathyroidism, osteomalacia & disease associatedwith Vitamin D defieciency.

PRINCIPLE

Determination of inorganic phosphorous according to the following reaction.

Phosphorous

Ammonium molybdate +sulfuric acid ------------------->phosphomolybdic complex

REAGENT COMPOSITION

INORGANIC PHOSPHOROUS REAGENT 4 x 25 mL / 4 x 50 mL

Sulfuric acid 210 mmol/L

Ammonium molybdate 650 mmol/L

INORGANIC PHOSPHOROUS STANDARD 1 x 4 mL

Phosphorous standard concentration 5 mg/dL

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 80C.

LINEARITY



NORMAL RANGE



Serum : 2.7 - 4.5 mg/dL

Urine : 400 - 1300 mg/24 hr.


Reagent and standard are ready to use.

PRECAUTION



NOTE: Use acid washed (50% HNO3) glasswares & tips.

SAMPLE

Serum / Plasma (free of haemolysis) / Urine (diluted 1/10 with distilled water).

INORGANIC PHOSPHOROUS4 x 25 mL, 4 x 50 mL

11012001, 11012002




Wavelength 340 nm

Temperature 370C


Linearity 15 mg/dL

Blank Reagent

Reaction Time 1 min







Standard - 20 µL -

Sample - - 20 µL

Mix & incubate at RT for 5 minutes. Read the absorbance of sample and standardagainst reagent blank.

CALCULATION


Phosphorous Conc. (mg/dL) = ------------------------------ x 5


BIBLIOGRAPHY

1. Tietz, N., Clinical Guide to Laboratory Tests, W.B. Saunders Company, Philad, 1983,384

2. Henry, R.J.; Clin. Chem., Harper & Row Publishers. New Yoork 19743. Thomas, L.; Labor and Diagnose, 2 Aufl. Med.Vert. Gem. Marburg 1979 4.

Taussky, H.H., Schorr, E.; J.Biol. Chem 202, 675 (1953)


44

INTENDED USE

This reagent is intended for in vitro quantitative determination of Iron in serum.

Iron Chromazurol method

Linearity- 500 µg/dL


In the blood iron is present in the hemoglobin of erythrocytes. Major function of ironin the body is the transportation of oxygen to the cells and cellular oxidation. Ironabsorbed in the small intestine and bound to a globulin in the plasma called transferrin.It is then transported to bone marrow where RBC generation take place.

Increased levels of iron in serum are seen in hemolytic aneamia, hepatitis and lead &iron poisoning.

Deceased levels are found in iron deficiency anemia, late pregnancy and cancer.

PRINCIPLE

Iron, bound to Transferrin in Fe(III) form, is released in an acidic medium and the Ferricions are reduced to Ferrous ions. The Fe (II) ions react with

Cromazurol B to form an intensely coloured complex. Intensity of the colour

formed is directly proportional to the amount of Iron present in the

sample.

REAGENT COMPOSITION

IRON REAGENT 2 x 50 mL

Acetate Buffer (pH 4.7) 0.2 mol/L

CTMA Bromide 0.7 mmol/L

Cromazurol B 2 mmol/L

IRON STANDARD 1 x 4 mL

Iron Standard (Concentration) 200 µg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 -80C.

LINEARITY


If the concentration is greater than linearity (500 µg/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum Iron (Males) : 59 - 158 µg/dL

(Females) : 37 - 145 µg/dL


Ready to use reagent.

PRECAUTION




SAMPLE

serum

IRON 2 x 50 mL

11022001




Wavelength 630 nm

Temperature 30oC



Blank Ragent Blank









Standard - 40 µL -

Sample - - 40 µL

Mix and incubate for 10 minutes at room temperature. Read the absorbance(A) ofstandard and sample against blank at 630 nm. The colour is stable for 1 hour whenprotected from light.

CALCULATION


Iron µg/dL = -------------------------------- x 200


BIBLIOGRAPHY

1. Weippl, G., et al: Blut.27.261 (1973)2. Garcic, A. ; Clin, chem, Acta 94,115 (1979)3. Tobacco, A., et al; Chem. Clin. Acta 114, 267 (1981)4. Teruzzi, A. , Torelli, G. ; 17th Meeting S.I. Bio.C.Clin.9,1080,communication (1985)

ADL/V.01/APR 2012

45

INTENDED USE

This reagent is intended for in vitro quantitative determination of Magnesium in serumor plasma.

- Xylidyl Blue with ATCS



Magnesium is the second most abundant intracellular cation of the human body afterpotassium, being essential in a great number of enzymatic and metabolic processes.

It is a co-factor of all the enzymatic reactions that involve ATP and found in themembranes that maintain the electrical excitability of muscular and nervous cells.

A low magnesium level is found in malabsorption syndrome, diuretics aminoglucosidetherapy, and hyperparathyroidism or diabetic acidosis.

Elevated concentration of magnesium is found in uremia, chronic renal failure,glomerulo nephritis, Addison’s disease or intensive anti acid therapy.

Clinical diagnosis is should not be made on a single test result; it should integrateclinical and other laboratory data.

PRINCIPLE

Magnesium reacts with xylidyl Blue to form a colored compound in alkaline solution.

The intensity of the color formed is proportional to the magnesium in the sample.

REAGENT COMPOSITION

MAGNESIUM REAGENT 4 x 25 mL

Xylidyl Blue 110mmol/L

Ethanolamine (pH 11.0) 1 mol/L

GEDTA 60 mmol/L

MAGNESIUM STANDARD 1 x 3 mL

Magnesium standard concentration 2 mg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-80C, protected from light.

LINEARITY



NORMAL RANGE



Serum : 1.8 – 2.6 mg/dL

CSF : 2.1 – 3.3 mg/dL

Urine : 73 - 122 mg/24 h



PRECAUTION

To avoid contamination, use clean laboratory wares. It is recommended to usedisposable tubes. Use clean, dry disposable pipette tips for dispending. Close reagentand standard bottles immediately after Use. Avoid direct exposure of reagent to light.

SAMPLE

Serum (free haemolysis) / Heparinized plasma (Do not use oxalates or EDTA asanticoagulant) / Urine (should be acidified to pH 3-4 with concentrated HCL thendilute sample 1/5 with distilled water and multiply the result by 5).

MAGNESIUM 4 x 25 mL

11019001




Wavelength 546 nm ( 520-550 nm)

Temperature 370C


Linearity 5 mg/dL

Blank Reagent







Reagent 1 1000 µL 1000 µL 1000 µL

Standard - 10 µL -

Sample - - 10 µL

Mix & incubate for 5 minutes at 370C. Read the absorbance of sample and standardagainst reagent blank.

CALCULATION

Magnesium Conc. (mg/dL) =


----------------------------- x Standard concentration


Unit Conversion

mg/dL x 0.4114 = mmol/L

mg/dL x 0.82 = mEq/L

BIBLIOGRAPHY

1. Farrel, E. C.; Magnesium.in Kaplan, A., et al.; Clin chem. The CV Mosby Co. St. Louis,Torento, Princeton 1984; 1064-69

2. Brutis, C. A., et al. TIETZ Text book of Clinical chemistry, 3 rd edition W. B Saunderscompany; 1999, P.1395-1457

3. Young, D. S.; Effects of disease on clinical lab tests, 4th edition, AACC Press,2001.


46

INTENDED USE

This reagent is intended for in vitro quantitative determination of total protein in urine.

- Pyrogallol Red method


- Sensitivity of 5 mg/dL


The presence of protein in urine is a very sensitive indicator of renal disorders. Thereare four ways by which increased amounts of protein can occur:increased glomerularpermeability; defective tubular re-absorption; increased plasma concentration of anabnormal, low molecular weight protein; and abnormal secretion of protein into theurinary tract.

Albuminuria, increased amounts of albumin in urine, has been recognized as an earlyindicator of renal damage in diabetes that can be reversed if detected and treated early.

PRINCIPLE

Proteins,in an acidic medium, combine with Pyrogallol Red and molybdate to form ablue purple coloured complex. The intensity of colour formed is directly proportionalto the amount of proteins present in the sample.

Proteins + Pyrogallol Red + Molybdate ----------> Blue purple coloured Complex

REAGENT COMPOSITION

MICROPROTEIN REAGENT 1 x 50 mL

Succinate buffer, pH 2.5 0.05 mmol/L

Sodium dodecylsulphate 0.07 mmol/L

Sodium molybdate 0.04 mmol/L

Pyrogallol-Red 0.06 mmol/L

MICROPROTEIN STANDARD 1 x 3 mL

Stabilized protein solution 100 mg/dL

Verified against NIST reference material.



LINEARITY

This reagent is linear up to 500 mg/dL with a sensitivity of 5mg/dL.


NORMAL RANGE



Urine < 24 mg/dL

Urine 24 hour < 120 mg /24 hour

CSF < 40 mg/dL



PRECAUTION



SAMPLE

Urine & CSF

Note: For haemolized or turbid samples it is suggested to carry out a sample blank:add 10 µL of sample to 1 mL of DI water and read the absorbance at 600 nm againstDI water.




Wavelength 600 nm

Temperature 370C

Blank Reagent

Standard concentration 100 mg/dL

Linearity 500 mg/dL

Incubation Time 5 minutes








Standard - 10 µL -

Sample - - 10 µL

Accurately mix and, after 5 minutes, read the absorbance of standard and sampleagainst blank at 600 nm. The colour is stable for 30 minutes.

CALCULATION

urine:

A sample

Proteins, mg/dL = -------------- x 100

A standard

Urine 24hrs:

A sample x1000 x T.V in litres

Proteins, mg/24 hrs = ---------------------------------

A standard

T.V = Total volume of 24 hr urine in milliitres

BIBLIOGRAPHY

Watnabe, N., et al.; Urinary protein as measured with a pyrogallol red-molibdatecomplex. Manually and Hitachi 726 automated analyzer. Clin. Chem. 32:1551-4 (1986)

MICROPROTEIN1 x 50 mL

11021001

47

INTENDED USE

This reagent is intended for in vitro quantitative determination of Total Iron BindingCapacity in serum.


Total iron-binding capacity (TIBC) is the measure of the ability of serum

proteins, principally transferrin, to bind iron. It is the maximumconcentration

of iron that the serum proteins can bind.

Increase in TIBC is found in Iron deficiency anemias and pregnancy.Decrease in TIBC is found in hypoproteinemia, hemolytic / pernicious /sickle cell anemias, inflammatory diseases and cirrhosis.

PRINCIPLE

TIBC is determined by addition of sufficient Fe(III) to saturate ironbinding sites on apotransferrin. The Fe3+ is removed by adsorption withbasic magnesium carbonate powder. After centrifugation bound ironremaining in supernatant is measured.

The difference between TIBC values and serum iron gives theconcentration of unsaturated transferrin(UIBC).

REAGENT COMPOSITION

TIBC REAGENT A 1 x 5 mL

Iron saturating solution 25 mg/dL

TIBC REAGENT B 50 x 50 mg

Magnesium Carbonate (Powder)


The sealed reagents are stable up to the expiry date stated on the label,when stored at 15-250C and protected from light.

LINEARITY

This reagent is linear up to 500µg/dL.

If the concentration is greater than linearity (500µg/dL), dilute thesample with normal saline and repeat the assay. Multiply the resultwith di lution factor.

EXPECTED VALUES

It is recommended that each laboratory establish its own referencevalues.


TIBC : 250 - 400 µg/dL

UIBC : 160 - 360 µg/dL



PRECAUTION

To avoid contamination, use clean laboratory wares. It is recommendedto use disposable tubes. Use clean, dry disposable pipette tips fordispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light.


SAMPLE

Serum (free from haemolysis)

TIBC 50 Test

11023001




Wavelength 630 nm

Temperature RT



Blank Reagent Blank






1. Serum Saturation procedure

Serum 500 µL

Reagent A 10 µL

Mix and leave at room temperature for 10 minutes; then add one aliquote of reagentB and shake; leave at room temperature for 15 minutes shaking at regular intervals (4times with vortex for 10-15 seconds). Centrifuge until you get a clear supernatant.

After centrifugation seperate the clear supernatent from the precipitate, and determinethe TIBC using iron reagent.

2. TIBC Determination


Iron Reagent 1000 µL 1000 µL 1000 µL


Iron Standard - 40 µL -

TIBC supernatent - - 40 µL

Mix and incubate for 10 minutes at room temperature. Read the absorbance(A) ofstandard and sample against blank at 630 nm. The colour is stable for 1 hour ifprotected from light

CALCULATION


TIBC µg/dL = --------------------------------- x 200


UIBC = TIBC - Iron level

ADL/V.01/APR 2012

48

INTENDED USE

This reagent is intended for in vitro quantitative determination of Total Protein inserum or plasma.

- Direct Biuret Method

- Linearity up to 15 gm/dL


Proteins form the major portion of dissolved substances in the plasma. They formthe basic structural components of the body. They constitute the enzymes presentin our body & also act as secondary source of energy. The other functions includedistribution of water, buffering, transport of various components, defense &coagulation of blood in our body.

Increased levels are found in dehydration & myeloma.

Decreased levels are found in liver disorders, Nephrotic syndrome, malnutrition &protein due to haemorrhage.

PRINCIPLE

Colorimetric determination of total protein based on the principle of the Biuret reaction(copper salt in an alkaline medium). Protein in plasma or serum sample forms a bluecolored complex when treated with cupric ions in alkaline solution. The intensity ofthe blue color is proportional to the protein concentration.

REAGENT COMPOSITION

TOTAL PROTEIN REAGENT 4 x 50 mL

Potassium iodide 6 mmol/L

Potassium sodium tartarate 21 mmol/L

Copper Sulphate 6 mmol/L


TOTAL PROTEIN STANDARD 1 x 3 mL

Total protein standard Concentration 6 gm/dL

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat room temperature and standard at 2- 80C.

LINEARITY

The procedure is linear up to 15 gm/dL. If the concentration is greater than linearity(15 gm/dL), dilute the sample with normal saline and repeat the assay. Multiply theresult with dilution factor.

NORMAL RANGE



Serum : 6.2 – 8.0 gm/dL



PRECAUTION



SAMPLE





Wavelength 546 nm

Temperature 370C

Standard Concentration 6 g/dL

Linearity 15 g/dL

Blank Reagent








Standard - 20 µL -

Sample - - 20 µL

Mix and incubate for 10 minutes at 370C. Measure the absorbance of standard andsample against reagent blank.

CALCULATION

Absorbacne of sample

Total Protein Conc. (g/dL) = ------------------------------ x 6


BIBLIOGRAPHY

Gomall, A.; J.Biol. Chem, 177 C (1949) 751

TOTAL PROTEIN4 x 50 mL

11013003

49

INTENDED USE

This reagent is intended for in vitro quantitative determination of Zinc in serum.


Zinc is needed for the body’s defensive (immune) system to properly work. It playsa role in cell division, cell growth, wound healing, and the break down ofcarbohydrates . Z inc is also needed for the senses of smell and taste. Z incsupplements in large amounts may cause diarrohea, abdominal cramps, andvomiting. Decreased levels are found in cirrhosis, lung carcinomas, sickle cellanemia, acute myocardial infarction, renal failure, corticosteroid and oralcontraceptive therapy .

Although zinc is an essential requirement for good health, excess zinc can be harmful

PRINCIPLE

Zinc in an alkaline medium reacts with Nitro-PAPS to form a coloured complex.Intensity of the colour is directly proportional to the amount of Zinc present in thesample.

REAGENT COMPOSITION

ZINC REAGENT 1 2 x 8 mL

Borate buffer(pH 8.2) 0.3 M

Sal ic i la ldox ime 12.5 mM

Dimethylgl ioxime 1.25 mM

ZINC REAGENT 2 2 x 2 mL

NITRO-PAPS 0.4mM

Preservatives

ZINC STANDARD 1 x 4mL

Z inc Standard (Concentration) 200 µg/dL


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-80C and protected from light.

LINEARITY


If the concentration is greater than linearity (1000 µg/dL), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

REFERENCE VALUES



Serum or Plasma : 70 - 115 µg/dL

Urine (24 hours) : 100 - 1000 µg/dL


Mix 4 parts of Reagent 1 with 1 part of Reagent 2.

PRECAUTION

To avoid contamination, use clean laboratory Wares. It is recommended to usedisposable tubes. Use clean, dry disposable pipette tips for dispensing. Close reagentand standard bottles immediately after use. Avoid direct exposure of reagent tolight.

SAMPLE

Serum or Plasma (free haemolysis). Use heparin as anticoagulant.

Urine (24 hours)

ZINC 2 x 10 mL

11024001




Wavelength 578 nm

Temperature 30oC



Blank Reagent Blank







WorkingReagent 1000 µL 1000 µL 1000 µL


Standard - 50 µL -

Sample - - 50 µL

Mix and incubate for 5 minutes at 30oC. Read the absorbance(A) of standard andsample against blank at 578 nm. The colour is stable for 30 minutes.

CALCULATION


Zinc µg/dL = ------------------------------- x 200


INTERFERENCE

Bilirubin uo to 20 mg/dL does not interfere.

BIBLIOGRAPHY

1. Pasquinelli, F.; Diagnostica e Tecniche di Laboratorio, (pag.:1103-1104) RossiniEditrice.(1984)

2. Testsuo Makino; Chimica Clinica Acta 197, 209-220(1991)3. Maringonia, A., Illuzi, R, ATB 1991 Abstract.

ADL/V.01/APR 2012

50

INTENDED USE

This reagent is intended for in vitro quantitative determination of Alpha1- AcidGlycoprotein in human serum

- Turbidimetric Immunoassay


- Ready to use reagents

- Multipoint calibration


Alpha-1-acid Glycoprotein is an acute-phase serum protein that is produced by theliver in response to inflammation and infection. AGP is useful in monitoring tumorrecurrence. Levels are also helpful in differentiating acute phase responses (elevatedlevels) from estrogen effects (normal or depressed levels). In addition, it is anexcellent protein in assessing in vivo hemolysis.

PRINCIPLE

The reagents containing polyclonal goat antihuman AGP when mixed with the serumsample containing AGP cause changes in absorbance, due to the development ofturbidity, which is directly proportional to the concentration of Alpha 1- AcidGlycoprotein in the sample.

REAGENT COMPOSITION

ALPHA 1- ACID GLYCOPROTEIN (AGP) R1 1 x 30 mL

Phosphate buffered saline (pH 7.43)

Polyethylene glycol (60 g/L)

Sodium azide (0.95 g/L)



Polyclonal goat anti-human Alpha 1- Acid Glycoprotein

(variable)


Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80C.

NORMAL RANGE



Male : 50 - 130 mg/dL


PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent bottles immediately after use. Avoid direct exposureof reagent to light.

SAMPLE

Use fresh serum. Dilute sample/control to 1/10 with saline.

GENERAL SYSTEM PARAMETERS FOR SEMI AUTO FOR FULLY AUTO

Mode of reaction End point End point



Temperature 37oC 37oC

Calibrator concentration As on vial label x Dilution factor

Linearity 300 mg/dL 300 mg/dL

Blank Reagent Blank Reagent Blank

Incubation time 5 min +5 min 5 min +5 min

Sample volume 5 µL 2.5 µL

Reagent 1 volume 500 µL 300 µL



CALIBRATION

Preparation of calibration curve:

Dilute the high concentrated calibrator 1/10 using saline and use this diluted calibratorfor the preparation of calibration curve.

Prepare the following calibrator dilution using normal saline as diluent. Multiply theconcentration of the AGP calibrator by the corresponding factor stated in the tablebelow to obtain the AGP concentration of each dilution.

Dilution 1 2 3 4 5 6

1/10dil.cal.(µL) - 10 10 25 50 100

Saline(µL) 100 150 70 75 50 -

Dil.factor 0 0.0625 0.125 0.25 0.5 1.0

LABORATORY PROCEDURE FOR FULLY AUTO

Blank Calibrator Sample/control

AGP R 1 300 µL 300 µL 300 µL

Dil.Calibrator - 2.5 µL -

Dil.Sample/control - - 2.5 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance (A1) at 340 nm.

AGP R 2 30 µL 30 µL 30 µL

Mix well and incubate for 5 minutes at 37°C. Measure the absorbance (A2) at 340nm.

Alternative Procedure for Semi autoanalyzer:


AGP R 1 500 µL 500 µL 500 µL

Dil.Calibrator - 5 µL -

Dil.Sample/control - - 5 µL

Mix and incubate for 5 minutes at 37°C.

AGP R 2 50 µL 50 µL 50 µL

Mix well and incubate for 5 minutes at 37°C. Measure the absorbance against thereagent blank at 340 nm.

CALCULATION

Multi point calibration

Calculate the Abs, plot a standard curve & read the concentration of controls &samples.

PERFORMANCE CHARACTERISTICS:

Measuring Range:- 4- 300 mg/dL.

If the concentration is greater than linearity (300 mg/dL), dilute the diluted(1/10)sample with normal saline and repeat the assay. Multiply the result with dilution factor.

Prozone Effect:- >600 mg/dL

Precision in CV%:-

Low Medium High

Intra - Run 4.66 1.14 2.45

Inter - Run 2.55

Accuracy in mg/dL:-

control Assigned value Measured value

level 1 44.3(35.5-53.2) 45.98

level 2 69.3(55.5-83.2) 72.3

level 3 89 (71.2 - 107) 94.1

INTERFERENCE

No interference for

Hemoglobin upto 1000 mg/dL

Na- citrate upto 1000 mg/dL

Heparin upto 50 mg/dL

Bilirubin upto 20 mg/dL

Triglyceride upto 2500 mg/dL

BIBLIOGRAPHY

1. Schmid,K. in FW Putman (Ed), The plasma protein,Vol 1, Second edition, AcademicPress, New York 2975, ppt 184-228

2. Johnson, A.M. et al., J. Clin. Invest., 48 (1969)22933. Dati,F. et al.,Lab.Med. 13 (1989)87

ALPHA 1-ACID GLYCOPROTEIN WITH CALIBRATOR1 x 30/1 x 3/1 mL

11822002


51

INTENDED USE

This reagent is intended for in vitro quantitative determination of Apolipoprotein A1in serum or plasma.

-Turbidimetric Immunoassay

-Linear up to 300 mg/dL

-Ready to use reagents

-Multipoint calibration


Apo A1 is the main protein component of HDL. Apo A1 activates lecithin cholesterolacyltransferase which catalyses the esterification of cholesterol this can then betransported to the liver, metabolized and excreted. People with atherosclerotic vascularchanges frequently exhibit decreased levels of Apo A1. Even if the concentrations ofapolipoprotein B are normal, a decreased ApoA1 level may be a risk factor foratherosclerosis. Decreased levels of ApoA1 also occur in dyslipoproteinemias, acutehepatic cirrhosis and insulin treated patients.

PRINCIPLE

Anti-human Apo A1 antisera when mixed with human serum containing Apo A1, reactto cause an absorbance change , which is measured by immunoturbidometricprinciple. The change in the absorbance can be interpolated in a calibration curveprepared with different known concentrations of calibrator.

REAGENT COMPOSITION

Apo A1 R1 1 x 30 mL


Polyethylene glycol 60 g/L

Detergent 0.1%

Sodium azide 0.95 g/L

Apo A1 R2 1 x 5 mL


Polyclonal goat anti-human Apo-A1(Variable)

Sodium azide 0.95g/L

Calibrator 1 x 1 mL



The sealed reagents are stable up to expiry date stated on the label, when stored at 2- 8oC.

REFERENCE RANGE

It is recommended that , each laboratory establish its own

reference values. The following values may be used as guidelines.

Men : 107 - 177 mg/dL

Women : 107 - 205 mg/dL

PRECAUTION

To avoid contamination, use clean laboratory wares; use clean dry disposable pipettetips for dispensing, close reagent bottle immediately after use . Avoid direct exposureof reagent to light.

SAMPLE

Use fresh serum. Dilute sample/control to 1 /10 with saline


Mode of reaction Fixed Time End point



Temperature 37o C 37o C



Blank DI Water Reagent Blank

Incubation time 5 min +5 min 5 min +5 minDelay 5 sec -

Delta 300 sec -





CALIBRATION


Reconstitute the Apo A1 calibrator with 1 mL of distilled water. The reconstitutedcalibrator is stable for 7 days at 2-8oC.

Dilute the high concentrated calibrator 1/10 using normal saline and use this dilutedcalibrator for the preparation of calibration curve.

Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the Apo A1 calibrator by the corresponding factors stated in the tablebelow to obtain the Apo A1 concentration of each dilution.


1/10dilCali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



Apo A1 R 1 240 µL 240 µL 240 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance (A1) at 340 nm.

Apo A1 R 2 40 µL 40 µL 40 µL


ALTERNATIVE PROCEDURE FOR SEMI AUTO ANALYZER:

Calibrator Sample/control

Apo A1 R 1 450 µL 450 µL

Dil.Calibrator 5 µL -

Dil.Sample/control - 5 µL

Apo A1 R 2 75 µL 75 µL

Mix well, measure absorbance A1 immediately after addition of ApoA1 R2 and takeabsorbance (A2) exactly after 300 sec at 340 nm.

CALCULATION

Multipoint calibration

Calculate Abs plot standard curve and read the concentration of controls andsamples.


Measuring Range:- 4 - 300 mg/dL.

If the concentration is greater than 300mg/dl dilute the diluted(1/10) sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

Prozone Effect:- >5500mg/dL

Precision in CV%:-

Low Medium High

Intra - Run 3.05 1.12 1.48

Inter - Run 1.63

Accuracy in mg/dL


evel 1 126(100-152) 137.62

level 2 88(70-106) 88.6

INTERFERENCE

No interference upto

Hemoglobin 1000mg/dL

Triglyceride 2500 mg/dL

Bilirubin 20mg /dL

BIBLIOGRAPHY

1. Tillett. W. S.et al: Serological reactions in pneumonia with a non-protein samaticfraction of pneumococcus. J.Exp.Med..52,561(1930)

2. Ziegenhagen G, Drahovshy D. klinishe Bedeutung des Creaktiven protein. Med klin1983;78:45-50.

3. Rifal. N.Tracy.R.P.Ridker, P.M.; Clinical efficacy of an Automated Highsensitivity C-Reactive protein Assay. Clin. chem.45:12.

Apo A1 WITH CALIBRATOR1 x 30/1 x 5/1 mL

11811001

52

INTENDED USE

This reagent is intended for in vitro quantitative determination of Apo B in serum.

-Turbidometric Immuno assay

-High Linearity of 330 mg/dL


-Multi point calibration


Apo B is the main protein component of LDL. It is necessary for the reaction with LDLreceptors in the liver and on cell walls and thus involved in transporting cholesterol,from the liver to the vessel cells.

Elevated levels of Apo-B are frequently found in atherosclerotic vascular changes andare a risk factor for atherosclerosis.

PRINCIPLE

The reagents containing polyclonal goat antihuman Apo-B antibodies when mixed withthe serum sample containing Apo-B cause changes in absorbance due to thedevelopment of turbidity, which is directly proportional to the concentration ofApo-B in the sample.

REAGENT COMPOSITION

Apo B R1 1 x 30 mL


Polyethyleneglycol 60 g/L

Detergent 0.1%

Sodium azide. 0.95 g/L

Apo B R2 1 x 5 mL

Polyclonal goat anti-human Apo-B(Variable)


Phosphate buffer pH 7.43

Calibrator 1 x 1 mL



The sealed reagents are stable upto the expiry date mentioned on the label when storedat 2-8oC.

REFERENCE RANGE

It is recommended that, each laboratory establishes its own reference values. Thefollowing values may be used as reference.

Men : 60 - 138 mg/dL

Women : 52 - 129 mg/dL

PRECAUTION

To avoid contamination, use clean laboratory wares, such as pipette tips and testtubes. Close reagent bottle immediately after use. Avoid direct exposure of reagentto light.

SAMPLE

Use fresh serum . Dilute sample/ control to 1/10 in saline

GENERAL SYSTEM PARAMETERS Semi Auto Fully Auto








Incubation time - 5 min +5 minDelay 5 sec -

Delta 300 sec -





CALIBRATION


Reconstitute the Apo B calibrator with 1 mL of distilled water. The reconstitutedcalibrator is stable for 7 days at 2-8oC. Dilute the high concentrated calibrator 1/10using saline and use this diluted calibrator for the preparation of calibration curve.

Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the Apo B calibrator by the corresponding factor stated in the tablebelow to obtain the Apo B concentration of each dilution.


1/10Dil.cal.(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



Apo B R 1 240 µL 240 µL 240 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance(A1) at 340 nm.

Apo B R 2 40 µL 40 µL 40 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance(A2)at 340 nm.

ALTERNATIVE PROCEDURE FOR SEMI AUTO ANALYZER


Apo B R 1 450 µL 450 µL


Dil.Sample/control - 15 µL

Apo B R 2 75 µL 75 µL

Mix well, measure absorbance immediately and after addition of Apo B R2 (A1)andtake absorbance exactly after 300 sec(A2) at 340 nm.

CALCULATION


Calculate abs, plot a standard curve & read the concentration of controls andsamples.



If the concentration is greater than 300 mg/dL , dilute thediluted(1/10) sample withnormal saline and repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 5.24 1.16 0.91

Inter - Run 1.02

Accuracy in mg/dL


Biorad level 1 84.4(67.8-101) 83.16

Biorad level 2 47.2(37.6-56.8) 47.39

IINTERFERENCE

No interference for



Bilirubin upto 20 mg /dL

BIBLIOGRAPHY

1. Tillett. W.S. et al: Serological reactions in Pneumonia a non-protein somatic fractionof pneumococcus. J. Exp.Med..52,561(1930)

2. Ziegenhagen G, Drahovshy D. klinishe Bedeutung des.Creaktiven protein. Med klin1983;78:45-50.

3. Rifal. N.Tracy.R.P.Ridker, P.M.clinical efficacy of anAutomated High sensitivity C-Reactive protein Assay. Clin. chem. 45:12.

Apo B WITH CALIBRATOR 1 x 30/1 x 5/1 mL

11812001

53

INTENDED USE

This reagent is intended for in vitro diagnostic quantitative determination of anti-Streptolysin O (ASO) in serum.

Linear up to 800 IU/mL

Single point calibration


Streptolysin O is a toxic immunogenic exoenzyme produced by beta-hemolyticstreptococcus group A, C and G. Measuring the antibodies ASO is useful for thediagnosis of rheumatoid fever, acute glomerulonephritis and streptococcal infections.Rheumatic fever is an inflammatory disease affecting connective tissue from severalparts of human body such as skin, heart joints etc., and acute glomerulonephritis isa renal infection that affects mainly renal glomerulus.

PRINCIPLE

The reagent ASO-Turbilatex agglutination assay is a quantitative turbidimetric assay formeasurement of ASO in human serum or plasma.

Latex particles coated with streptolysin O are agglutinated when mixed with samplescontaining ASO. The agglutination causes an absorbance change, dependent upon theASO contents of the patient sample that can be quantified by comparison from acalibrator of known ASO concentration.

REAGENT COMPOSITION

ASO TURBILATEX R1 1 x 45 mL

Diluent

Tris buffer 20 mmol/L


ASO TURBILATEX R2 1 x 5mL

ASO-Latex:

Suspension of latex particles coated with

streptolysin O, (pH 10.0)


ASO CALIBRATOR 1 x 1 mL

Calibrator concentration is stated on the vial label.

PRECAUTIONS: Components from human origin have been tested and found to benegative for the presence of HbsAg, and HCV and of antibody to HIV (1/2). Howeverhandle cautiously as potentially infectious.

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-8oC.

NORMAL RANGE

It is recommended that each laboratory establish its own reference value. The followingvalue may be used as guideline.

Serum

up to 200 IU/mL(adults)

up to 100 IU/mL (children below 5 years)


ASO calibrator:Reconstitute the calibrator with 1mL distilled water. Reconstitutedcalibrator is stable for 30 days at 2-8oC.

PRECAUTION


SAMPLE

Fresh serum. (Do not use hemolyzed or lipemic serum)

GENERAL SYSTEM PARAMETERS

Mode of reaction Fixed time



Temperature 370C

Calibrator concentration As on vial label

Linearity 800 IU/mL

Blank DI water

Delay time 5 sec

Interval 120 sec




Cuvette 1 cm path length



ASO reagent 1 900 µL 900 µL

Calibrator 10 µL -

Sample/control - 10 µL

ASO reagent 2 100 µL 100 µL

Mix and read the absorbance immediately (A1) and after 2 minutes (A2) of sampleaddition at 546 nm.

CALCULATION

(A2-A1) sample x calibrator concentration

ASO con. in IU/mL = -------------------------------------------------------------

(A2-A1) calibrator

PERFORMANCE CHARACTERISTICS

Measuring range: 20-800 IU/mL

The reagent is linear up to 800 IU/mL.


1. Detection limit: Values less than 20 IU/mL gives non reproducible results2. Prozone effect: No prozone effect was detected up to 2000 IU/mL.

INTERFERENCES

No interference

Rheumatoid factors : up to 300 IU/mL

Bilirubin : up to 20mg/dL

Lipemia : up to 10g/L.

BIBLIOGRAPHY

1. Haffejee, Quarterly Journal of Medicine 1992, New Series 84; 305: 641-6582. Alouf et al Biochemie. 1973; 56-61.

ASO TURBILATEX WITH CALIBRATOR1X45 mL/1X5 mL/1 mL

11801001

54

INTENDED USE

This reagent is intended for in vitro quantitative determination of Anti Streptolysin –O (ASO).

-Latex enhanced immunoturbidimetry


-No sample dilution required

-Linear up to 800 IU/mL


ß - hemolytic streptococcus bacteria especially group A, C and G, produce an exotoxinknown as Streptolysin-O. People infected with this bacterium produce an antibodyknown as Anti Streptolysin-O (ASO). Measuring the levels of ASO is effective fordiagnosing, judging the progress of medical treatment and assessing the recovery fromdiseases like rheumatic fever, acute glomerulonephritis and tonsillitis.

PRINCIPLE

When an antigen-antibody reaction occurs between ASO in the sample andstreptolysin-O which has been sensitized to latex particles, agglutination occurs. Thisagglutination results in change of absorbance and is proportional to the quantity ofASO in the sample. The concentration can be determined by comparison with acalibrator of known ASO concentration

REAGENT COMPOSITION

ASO R 1 1 x 24 mL, 2 x 24 mL

Glycine buffer solution

ASO R2 1 x 8 mL, 2 x 8 mL

ASO Latex suspension particles coated with Streptolysin - O

CALIBRATOR 1 x 1 mL, 1 x 2 mL

ASO calibrator concentration as on vial label



NORMAL RANGE



Serum

Adults : 200 IU/mL

children < 5 years : 100 IU/mL

PRECAUTION

To avoid contamination use clean laboratory wares. Use clean dry disposable pipettetips for dispensing. Close reagent bottles immediately after use. Avoid direct exposureof reagent to light.

SAMPLE

Fresh serum / plasma

GENERAL SYSTEM PARAMETER SEMI AUTO FULLY AUTO

Mode of Reaction Fixed time End point


Wavelength 578 nm 570/800 nm


Calibrator Concentration As on vial label As on vial label

Linearity 800 IU/mL 800 IU/mL

Blank DI water Reagent blank

Delay time 5 sec -

Interval 240 sec -







ASO R1 210 µL 210 µL 210 µL


Sample/control - - 3 µL

Incubate for 5 minutes at 370C.

ASO R2 70 µL 70 µL 70 µL

Mix and read the absorbance immediately (A1), and after 4 minutes (A2) at 570/800nm.

ALTERNATIVE PROCEDURE FOR SEMI AUTO


ASO R1 450 µL 450 µL

Calibrator 5 µL -


ASO R2 150 µL 150 µL

Mix and read the absorbance immediately (A1), and after 4 minutes (A2) at 578 nm.

CALCULATION

(A2-A1) sample

ASO Conc. in IU/mL = ------------------------- x Calibrator Conc.

(A2-A1) calibrator


Measuring Range:- 20 – 800 IU/mL.

If the concentration is greater than 800 IU/mL, dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.

Prozone Effect:- > 5600 IU/mL

Precision in CV%:-

Medium High

Intra - Run 5.0 3.0

Inter - Run 8 5

Accuracy in IU/mL


level 1 121(96.9-145) 105.7

level 2 198(158-237) 187.4

level 3 284(227-341) 247

INTERFERENCES

Do not interfere for



Intrafat up to 5000 mg/dL

BIBLIOGRAPHY

1. Galuin, J.P. et al.: Particle enhanced photometric immune assay system, Clin. LabAssays (pap .Annu.clin.Lab.Assays Conf.)

2. Singer J.M. et al. The latex fixation test Application to the serologic diagnosis orrheumatoid arthritis, Amer J.Med 21, 888(1956)

ASO LEIT WITH CALIBRATOR 1 x 24 /1 x 8 /1 mL, 2 x 24 /2 x 8 /2 mL

11807004, 11807005

55

INTENDED USE

This reagent is intended for in vitro quantitative determination of complement C3 inhuman serum.






complement C3 is the central point of the classic and alternative complement pathway.

Complement testing help to diagnose the cause of recurrent microbial infections,angioedema, or inflammation. It may be used to help diagnose and to monitor theactivity of acute or chronic autoimmune diseases such as Systemic LupusErythematosus (SLE).

Decreased levels of C3 are significant in autoimmune disease, immune infectionswith pyrogenic bacteria, bacteremia, neonatal respiratory distress syndrome andcongenital deficiencies.C3 behaves as an acute phase protein hence increased levelsmay found in acute inflammatory reactions.

PRINCIPLE

The reagents containing polyclonal goat antihuman C3 when mixed with the serumsample containing C3 cause changes in absorbance, due to the development ofturbidity, which is directly proportional to the concentration of C3 in the sample.

REAGENT COMPOSITION

C3 R1 1 x 30 mL




C3 R2 1 x 5 mL


Polyclonal goat anti-human C3C (variable)


Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C

NORMAL RANGE



Serum 75-135 mg/dL

PRECAUTION


SAMPLE

Use fresh serum. Dilute sample/control to 1/10 with saline. If the test cannot be carriedout on the same day, the serum may be stored at 2-80 C for 48 hours.





Temperature 37 oC 37 oC




Incubation time 5 min +5 min 5 min +5 min





calibration curve spline

CALIBRATION

PREPARATION OF CALIBRATION CURVE

Dilute the calibrator to 1/10 using normal saline and use this diluted calibrator forthe preparation of the calibration curve. Prepare the following calibrator dilution usingNaCl as diluent. Multiply the concentration of the C3 calibrator by the correspondingfactors stated in the table below to obtain the C3 concentration of each dilution.


1/10 Cali. (µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

LABORATORY PROCEDURE FOR FULLY AUTO ANALYZER

Blank Calibrator Sample/Control

C3 R 1 200 µL 200 µL 200 µL

Dil. Calibrator - 3 µL -



C3 R 2 30 µL 30 µL 30µL



Blank calibrator sample /control

C3 R1 450 µL 450 µL 450 µL

Dil calib - 5µL -

Dil sample/control - - 5µL

Mix and incubate for 5 minutes at 37oC.

C3 R2 75 µL 75 µL 75 µL

Mix and incubate for 5 minutes at 37oC. Measure the absorbance against reagentblank at 340 nm.

CALCULATION

Multipoint calibration.

Calculate the Abs. Plot a standard curve and read concentration of controls andsamples.


Measuring Range:- 20 –400 mg/dL.

If the concentration is greater than linearity(400 mg/dL), dilute the diluted(1/10)sample with normal saline and repeat the assay. Multiply the result with dilutionfactor.


Precision in CV%:-

Low Medium High

Intra - Run 2.82 3.43 3.28

Inter - Run 3.71 2.56

Accuracy in mg/dl:-


level 1 80.2(64.1-96.2) 78.57

level 2 166(133-199) 173.2

level 3 254(203-305) 249.7

INTERFERENCE

No interference for


Na-citrate upto1000 mg/dL

Heparin upto50 mg/dL


Triglyceride upto2500 mg/dL

BIBLIOGRAPHY

1. Dati, F. et al., Lab. Med.13, 87 (1989)2. Muller-Eberhard, H.H., Ann. Rev.Biochem.44, 697(1975)3. Lachmann, P.J., Hobart, M.J. and Ashton, W.P. (1973)

C3 WITH CALIBRATOR1 x 30/1 x 5/1 mL

11819001


56

INTENDED USE







C4 is a constituent of C3 convertase & C5 convertase.

Decreased levels are found in hereditary angioneurotic odema, immune complexdisease and congenital deficiencies.

PRINCIPLE


REAGENT COMPOSITION

C4 R1 1 x 30 mL

Phosphate buffered saline (pH7.43)



C4 R2 1 x 5 mL




CALIBRATOR 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C.

NORMAL RANGE



Serum : 9 - 36 mg/dL

PRECAUTION


SAMPLE

Use fresh serum. Dilute the sample/control to 1/10 with saline. If the test cannot becarried out on the same day, the serum may be stored at 2-80 C for 48 hours.









Incubation time 5 min +5 min 5 min +5 minSample volume 5 µL 3µL




calibration curve spline

CALIBRATION


Dilute the high concentration calibrator to 1/10 with normal saline and use thisdiluted calibrator for the preparation of calibration curve. Prepare the followingcalibrator dilution using NaCl as diluent. Multiply the concentration of the C4calibrator by the corresponding factors stated in the table below to obtain the C4concentration of each dilution.


1/10 dil.Cali. (µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



C4 R 1 200 µL 200 µL 200 µL




C4 R 2 30 µL 30 µL 30 µLMix and incubate for 5 minutes at 37°C. Measure the absorbance (A2) at 340 nm.



C4 R 1 450 µL 450 µL 450 µL




C4 R 2 75 µL 75 µL 75 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance against reagentblank at 340 nm.

CALCULATION


Calculate the Abs, plot a standard curve and read the concentration of controlsand samples.



If the concentration is greater than linearity (80 mg/dL), dilute thediluted(1/10) samplewith normal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 4.54 2.18 3.96

Inter - Run 4.17 3.08

Accuracy in mg/dL


level 1 12.7(10.1-15.2) 12.11

level 2 28.3(22.7-34.0) 27.4

level 3 41.8(33.5-50.2) 39.9

INTERFERENCE

No interference for


Na-citrate upto1000 mg/dL


Turbidity upto 5%


Triglyceride upto2500 mg/dL

BIBLIOGRAPHY

1. Dati, F. et al., Lab. Med.13, 87 (1989)2. Muller-Eberhard, H.H., Ann. Rev.Biochem.44, 697(1975)

C4 WITH CALIBRATOR1 x 30/1 x 5/1 mL

11820001


57

INTENDED USE

This reagent is intended for in vitro quantitative determination of Ceruloplasmin inserum.

-Turbidimetric Immuno assay


-No sample dilution needed




Ceruloplasmin is a copper oxidase enzyme, important in regulating the ionic state ofiron and other metallic ions. Levels are decreased in hepatolenticular degeneration orWilson's disease and Menke's kinky hair syndrome. Levels are elevated by the acutephase response and particularly by estrogens.

PRINCIPLE

The reagents containing polyclonal goat antihuman ceruloplasmin when mixed withthe serum sample containing ceruloplasmin cause changes in absorbance due tothe development of turbidity, which is directly proportional to the concentrationof Ceruloplasmin in the sample.

REAGENT COMPOSITION

Ceruloplasmin - R1 1 x 30 mL






Polyclonal goat antihuman Ceruloplasmin


Calibrator 1 x 1 mL



The sealed reagents are stable upto the expiry date mentioned on the label when storedat 2-8oC. Do not freeze.

REFERENCE RANGE

It is recommended that, each laboratory should establishes its own reference values.The following value may be used as a reference.

serum : 22 - 61 mg/dL

PRECAUTION


SAMPLE

Fresh serum.

GENERAL SYSTEM PARAMETERS Semi Auto Fully Auto






Linearity 100 mg/dl 100 mg/dl


Incubation time 5 min 5 min + 5min

Delay 5 sec -

Delta 300 sec -





CALIBRATION


Prepare the following calibrator dilution using normal saline as diluent. Multiply theconcentration of the Ceruloplasmin calibrator by the corresponding factors stated inthe table below to obtain the Ceruloplasmin concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0


Blank calib Sample/control

Ceruloplasmin R1 300 µL 300 µL 300 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance(A 1) at 340 nm.

Ceruloplasmin R2 50 µL 50 µL 50 µL




Ceruloplasmin R 1 450 µL 450 µL

Dil.calibrator 5 µL -


Ceruloplasmin R 2 75 µL 75 µL

Mix well, measure absorbance (A1)immediately after addition of Ceruloplasmin R2 andtake absorbance exactly after 300 sec (A2) at 340 nm.

CALCULATION





If the concentration is greater than 100 mg/dL , dilute the diluted(1/10)sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

Prozone Effect:- > 400 mg/dL

Precision in CV%:-

Low Medium High

Intra - Run 6.31 2.07 2.20

Inter - Run 3.91

Accuracy in mg/dL


level 1 18.7(14.9-22.4) 20.98

level 2 35.8(28.7-43.0) 38.3

level 3 50.2(40.1-60.2) 48.57

Interference:-

No interference for


Na - citrate upto 1000 mg/dL



Triglycerides upto 2500 mg/dL

BIBLIOGRAPHY

Poulik , M.D and Kleiss M.L in “The plasma proteins”, F.W. Putman, (ed.) Vol. 2Second edition, Academic press, New York, PP 52-108

CERULOPLASMIN WITH CALIBRATOR 1 x 30/1 x5 /1 mL

11813001


58

INTENDED USE

This reagent is intended for in vitro diagnostic quantitative determination of C-reactiveprotein (CRP) in serum.

Linear up to 150 mg/L



CRP (C – reactive Protein) is a cytokine - induced, acute phase protein that increasesin concentration as a result of inflammation. CRP levels in the body has been used asa marker or indicator of infections and inflammation. The assay of CRP is moresensitive than the erythrocyte sedimentation rate (ESR) and leukocyte count. The CRPlevels rise and return to reference ranges more rapidly after the disease has subsided.

PRINCIPLE

This is a quantitative turbidimetric immuno assay for the measurement of CRP inhuman serum. CRP in the samples binds to specific anti-CRP antibodies, which havebeen adsorbed to latex particles and agglutinates. The agglutination is proportionalto the quantity of CRP in the sample. The actual concentration is then determined byinterpolation from a calibration curve prepared from calibrators of knownconcentrations.

REAGENT COMPOSITION

CRP TURBILATEX R1 1x45 mL

Diluent

Tris buffer 20 mmol/L


CRP TURBILATEX R2 1x5 mL

CRP-Latex:


anti human CRP


CRP CALIBRATOR 1x1 mLCalibrator concentration is stated on the vial label.


STORAGE & STABILITY


NORMAL RANGE


Serum : up to 6 mg/L


CRP calibrator: Reconstitute the calibrator with 1mL of distilled water.Reconstitutedcalibrator is stable for 30 days at 2-80C.

PRECAUTION


SAMPLE



Mode of reaction Fixed time



Temperature 37oC

Calibrator concentration As on vial label

Linearity 150 mg/L

Blank DI water

Delay time 5 sec

Interval 120 sec

Sample volume 5 µL





Calibrator Sample /control

CRP R1 900 µL 900 µL

Calibrator 5µL ---

Sample --- 5 µL

CRP R2 100 µL 100 µL

Mix and read the absorbance immediately (A1) and after 2 minutes (A2) of sampleaddition.

CALCULATION

(A2-A1) sample x calibrator concentration

CRP Conc. in mg/L = -------------------------------------------------------

(A2-A1) calibrator


1. Linearity : The reagent is linear up to 150 mg/L. If the concentration is greater thanlinearity, dilute the sample with normal saline and repeat the assay. Multiply theresult with dilution factor.

2. Detection limit: Values less than 2mg/L gives non reproducible results3. Prozone effect: No prozone effect was detected up to 800 mg/L.

INTERFERENCES

No interference

Rheumatoid factors : up to 300 IU/mL

Bilirubin : up to 20 mg/dL

Lipemia : up to 10 g/L.

BIBLIOGRAPHY

1. Haffejee, Quarterly Journal of Medicine 1992, New Series 84; 305: 641-6582. Alouf et al Biochemie. 1973; 56-61.

CRP TURBILATEX WITH CALIBRATOR 1X45 mL/1X5mL/1X1mL

11802001

59

INTENDED USE

This reagent is intended for in vitro quantitative determination of C-reactive proteinin human serum or plasma by immunoturbidimetry.


-Linear up to 200 mg/L


-No sample dilution needed


CRP (C – Reactive Protein) is a cytokine - induced, acute phase protein that increasesin concentration as a result of inflammation. CRP levels in the body has been used asa marker or indicator of infections and inflammation. The assay of CRP is moresensitive than the erythrocyte sedimentation rate (ESR) and leukocyte count. The CRPlevels rise and return to reference ranges more rapidly after the disease has subsided.

PRINCIPLE

This is a latex enhanced turbidimetric immuno assay. CRP in the samples binds tospecific anti-CRP antibodies, which have been adsorbed to latex particles andagglutinates. The agglutination is proportional to the quantity of CRP in the sample.The actual concentration is then determined by interpolation from a calibration curveprepared from calibrators of known concentrations.

REAGENT COMPOSITION

CRP R1 1 x 24 mL, 2 x 24 mL

Glycine buffer

CRP R2 1 x 8 mL, 2 x 8 mL

Latex suspension coated with anti-CRP antibodies. (rabbit polyclonal antibody)

CALIBRATOR 1 x 2 mL, 1 x 2 mL

CRP calibrator concentration as on vial label



NORMAL RANGE

It is recommended that each laboratory should establish its own reference values.The following value may be used as a guide line.

Serum up to 6 mg/L

PRECAUTION


The reagent should be used according to this pack insert. If used otherwise,appropriate performance is not guaranteed.

SAMPLE

Fresh serum (Do not use hemolized or lipemic serum)


Mode of Reaction Fixed Time End point


Wavelength 578 nm 570/ 800nm


No.of calib. 6 6


Linearity 200 mg/L 200 mg/L

Blank DI Water Reagent blank

Delay 5 sec --

Interval 120 sec --




Cuvette 1cm light path 1cm light path

CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the CRP calibrator by the corresponding factor stated in the tablebelow to obtain the CRP concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0


Blank calibrator Sample/Control

CRP R 1 210 µL 210 µL 210 µL




CRP R 2 70 µL 70 µL 70 µL

Mix and measure the absorbance immediately (A1) and after 2 minutes (A2) at 570/800nm.


calibrator Sample/Control

CRP R 1 450 µL 450 µL

Dil. Calibrator 5 µL -


CRP R 2 150 µL 150 µL

Mix and measure the absorbance immediately (A1) and after 2 minutes (A2) at 578nm.

CALCULATION




Measuring Range:- 1 –200 mg/L.

If the concentration is greater than 200 mg/L, dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.

Prozone Effect:- >1000 mg/L

Precision in CV%:-

Low Medium High

Intra - Run 7.0 5.0 3.0

Inter - Run 10 8 5

Accuracy in mg/L


level 1 5.85(4.68-7.02) 5.05

level 2 27.3(21.9-32.8) 26.8

level 3 51.9(41.5-62.2) 49.6

INTERFERENCE

No interference for

Hemoglobin 500 mg/dL

Intrafat 500 mg/dL

Bilirubin 30 mg/dL

RF 500 IU/mL

BIBLIOGRAPHY

1. Tillett.W.S..et al: Serological reactions in pneumonia with a non protein somaticfraction of pneumococcus.J.Exp.Med..52,561(1930).

2. Zeigenhagen G,Drahovshy D.Klinishe Bedeutung des C-reaktiven protein.Med klin1983;78:45-50.

3. Rifal.N.Tracy.R.P.Ridker,P.M.Clinical efficacy of an Automated High sensitivity C-Reactive protein Assay: Clin chem. 45-12.

CRP LEIT WITH CALIBRATOR1 x 24 / 1 x 8 / 2 mL, 2 x 24 / 2 x 8 / 2 mL

11808004, 11808005

60

INTENDED USE

This reagent is intended for in vitro quantitative determination of C-reactive protein(CRP) in serum.

-Latex Enhanced Immuno Turbidimetric assay

-Sensitivity of 0.13 mg/L

-Linearity up to 10 mg/L

-Ready to use reagent


CRP is an acute phase protein produced by liver. It’s level will rise in response toinflammations and infections.

Inflammation of arteries is a risk factor for cardiovascular disease. It is linked to anincreased risk of heart disease, heart attack, stroke and peripheral arterial disease. CRPultra/HS CRP is the strongest predictor of cardiac risk. The value more than 10 mg/Lcannot be considered for cardiac risk factors.

Routinely available CRP methods are to determine infections or chronic inflammatorydisease where CRP concentration is above 10 mg/L. These methods are with limitedsensitivity hence it cannot precisely measure CRP concentrations below 10 mg/L.

Studies have shown that measuring CRP with improved methodology of high sensitiveassay can identify the risk level of CVD in apparently healthy people. Relatively highlevels of hs CRP in healthy individuals are predictive of the future risk of heart diseaseeven when cholesterol levels are within the acceptable range.

PRINCIPLE

This is a latex-enhanced turbidimetric invitro immuno assay. CRP in the sample bindsto specific anti-CRP antibodies, which had been adsorbed to latex particles andagglutinates. The agglutination is detected as an absorbance change. The magnitudeof the change is proportional to the concentration of CRP in the sample. The actualconcentration is then detected by interpolation from a calibration curve prepared fromcalibrators of known concentration.

REAGENT COMPOSITION

CRP Ultra R1 1 x 18 mL

Glycine buffer


Latex suspension coated with anti-CRP antibodies. (Rabbit polyclonal antibody)

CALIBRATOR 1 x 2 mL

Ready to use CRP calibrator, the concentration is as on the vial label

PRECAUTIONS: Components from human origin have been tested and found to benegative for the presence of HBsAg, and HCV and of antibody to HIV (1/2). However,handle cautiously as potentially infectious.


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-80C. Stability in the instrument is at least 4 weeks if contamination is avoided.Do not freeze.

REFERENCE RANGE



Less than 1 mg/L = Low risk for CVD

1.0-2.9 mg/L = Intermediate Risk for CVD

Greater than 3 mg/L = High Risk for CVD


The Reagent1 & Reagent 2 are ready to use.

Calibrator :Ready to use

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent bottles immediately after use.

Avoid direct exposure of reagent to light. Do not blow into the reagent bottles.

SAMPLE


GENERAL SYSTEM PARAMETER FULLY AUTO SEMI AUTO

Mode of Reaction End point Fixed time


Wavelength 570/800nm 578 nm


No.of standards 6 6

Calibrator Concentration As on vial label X Dilution Factor

Linearity 10mg/L 10 mg/L

Blank Reagent Blank DI water

Delay time - 5 sec

Interval - 120 sec




CALIBRATION


Prepare the following calibrator dilution using normal saline as diluent. Multiply theconcentration of the CRP ultra calibrator by the corresponding factors stated in thetable below to obtain the CRP ultra concentration of each dilution.


Calib.(µL) - 10 10 20 50 100

Saline(µL) 100 150 70 60 50 -

Dil. Factor 0 0.0625 0.125 0.25 0.5 1.0

LABORATORY PROCEDURE FOR SEMI AUTO ANALYSER


CRP ultra R 1 400 µL 400 µL

|Dil. Calibrator 10 µL -


CRP ultra R 2 200 µL 200 µL

Mix and read absorbance immediately(A1), and after 2 minutes(A2)of the sampleaddition at 578 nm.

LABORATORY PROCEDURE FOR FULLY AUTO ANALYSER


CRP ultra R 1 200 µL 200 µL 200 µL



Mix and incubate for 5 minutes at 370C

CRP ultra R 2 100 µL 100 µL 100 µL

Mix and read absorbance immediately(A1), and after 2 minutes(A2)of the R2 additionat 570/800 nm.

CALCULATION


Calculate the Abs , plot a standard curve & read the concentration of controls &samples.


Measuring Range:- 0.13 – 10 mg/L.


Precision in CV%:-

Low Medium High

Intra - Run 7 5 3

Inter - Run 10 8 5

Interference:-




Conjugated Bilirubin up to 30 mg/dL

Intra Fat up to 500 mg/dL

Rheumatoid Factor up to 500 IU/mL

BIBLIOGRAPHY

1. Claus, D. R; Osmand, A.P.; Gewurz, H. Radioimmunoassay of human C-reactiveprotein and levels in normal sera. J .Lab. Clin Med 1976;87: 120-128

2. Wasunna, A, Whitelaw, A. Gallimore, R. Hawkins, P.N. Pepys, M. B. C-reactive proteinand bacterial infection in preterm infants. Eur J Pediatr 1990; 149: 424-427

CRP ULTRA WITH CALIBRATOR 1x18 / 1x9 / 2 mL

11808006

61

INTENDED USE

This reagent is intended for in vitro quantitative determination of Cystatin C in humanserum.

-Latex enhanced Immunoturbidimetry





Cystatin C is a low molecular weight (13 Da) cytoplasmic protein, functioning as aninhibitor of various cystein protease in the blood stream. Cystatin C has a stableproduction rate and is removed from the blood circulation by glomerular filtration.In healthy individuals Cystatin C is completely reaborsorbed and degraded in thetubules but in subject with renal disorders its level in blood may be raised as high as2 to 5 times the normal values. Cystatin C is superior to serum creatinine as a markerof glomerular filtration Rate.

PRINCIPLE

Cystatin C in the test sample binds to the specific polyclonal rabbit anti-Cystatin Cantibody, which has been adsorbed to latex particle and agglutinates. The agglutinationis detected as absorbance change at 546 nm. The magnitude of change is proportionalto the quantity of Cystatin C in the sample and its concentration is determined byinterpolation from a calibration curve prepared from calibrators of knownconcentration.

REAGENT COMPOSITION

Cystatin C R1 1 x 25 mL

Tris buffer 1.2% (100 mM)

pH 8.5+0.3


Polystyrene latex particle coated with polyclonal anti Cystatin C antibody (rabbit)

CALIBRATOR 1 x 2 mL

Cystatin C calibrator concentration as mentioned on the vial label.



NORMAL RANGE



Individuals up to 50 years : 0.55 – 1.15 mg/L

Individuals above 50 years : 0.63 – 1.44 mg/L

PREPARATION OF REAGENT

Reagent 1 and Reagent 2 are ready to use.Cystatin –C calibrator: Ready to use Liquid stable.Unopened vials are stable till expiration date mentioned on the vials. Opened vials arestable for 4 weeks when stored at 2-80C.

PRECAUTIONS:To avoid contamination, use clean laboratory wares, such as pipette tips and testtubes. Close reagent bottle immediately after use. Avoid direct exposure of reagent tolight.

SAMPLE

Required sample material is human serum or EDTA/ Heparinized plasma. It isrecommended to analyze the sample as fresh as possible.


Mode of Reaction Fixed time End point


Wavelength 546 nm 546 nm/ 800nm


No of calib 6 6Calibrator Conc. As on vial label x Dilution factorsLinearity 10 mg/L 10 mg/LBlank DI Water Reagent blank

Delay Time 5 sec --

Interval 300 sec --Sample volume 5 µL 3 µLReagent 1 volume 500 µL 200 µL

Reagent 2 volume 100 µL 40 µLCuvette 1 cm light path 1 cm light path

Dilution of calibrator for calibration curve:

Calibration Curve (range between 0-10 mg/L). Prepare the following calibrator dilutionusing normal saline as diluent. Multiply the concentration of the Cystatin C calibratorby the corresponding factor stated in the table below to obtain the Cystatin Cconcentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 200 190 70 60 50 -

Dil. factor 0 0.05 0.125 0.25 0.5 1.0



Cystatin R1 200 µL 200 µL 200 µL




Cystatin R2 40 µL 40 µL 40 µL

Mix and read the absorbance immediately (A1), and after 5 minutes (A2) at 546/800nm.



Cystatin R1 500 µL 500 µL



Cystatin R2 100 µL 100 µL

Mix and read the absorbance immediately (A1), and after 5 minutes (A2) at 546nm.

CALCULATION


Calculate the abs, plot a standard curve & read the concentration of controls &samples.


Measuring Range:- 0.1 - 10 mg/L.

If the concentration is greater than 10 mg/L, dilute the sample with normal saline andrepeat the assay .Multiply the result with dilution factor.


Precision :

MEAN INTRA RUN INTER RUN nVALUE CV(%) CV(%)

Low human serum pool 0.77 2.16 2.54 20

High human serum pool 5.94 0.67 1.45 20

Medium human serum pool 1.45 1.58 1.95 20




Accuracy in mg/L


level 1 0.484(0.387-0.581) 0.438

level 2 0.572(0.457-0.686) 0.530

level 3 0.649(0.519-0.779) 0.614

IINTERFERENCE



Intrafat 1400 mg/dL

Bilirubin 25 mg/dL

BIBLIOGRAPHY

1. Cystatin-C as a marker of GFR - history, indications and future research; Clin.Biochem.38:1, 2005

2. Serum Cystatin -C is superior to serum creatinine as a marker of kidney function:a meta analysis. Am. J. Kidney Desease. 40, 221, 2002.

CYSTATIN-C WITH CALIBRATOR 1 x 25/1 x 5/2 mL

11810001

62

INTENDED USE

The reagent is intended for in vitro quantitative determination of Ferritin in serum

-Latex Enhanced Immunoturbidimetry

-High Linearity of 1000 ng/mL

-No sample dilution




Ferritin is an iron-containing protein. It is mainly found in liver and spleen, where itsfunction is to store and release iron in the body. It is also found in small amounts inhuman serum. The serum levels tend to increase due to hepatitis and malignanttumors. The measurement of ferritin is useful in diagnosis, treatment, assessment ofdisease progression and post operative prognosis of abnormal iron metabolism andiron deficiency anaemia.

PRINCIPLE

Latex particles coated with anti-ferritin antibody are agglutinated when mixed withsamples containing Ferritin. The agglutination causes an absorbance change whichdepends on the Ferritin concentration in the sample, this can be interpolated using acalibration curve prepared from calibrators of different concentrations.

REAGENT COMPOSITION

Ferritin - R1 1 x 30 mL

Glycine buffer


Suspension of latex particle bound to anti-ferritin antibodies

Calibrator 1 x 1 ml




NORMAL RANGE

It is recommended that, each laboratory should establish its own reference values. Thefollowing value may be used as a guide line.

Male : 30 - 220 ng/mL

Females : 20 - 110 ng/mL

PRECAUTION

To avoid contamination, use clean laboratory wares, use clean dry disposable pipettetips for dispensing, close reagent bottle immediately after use. Avoid direct exposureof reagent to light.

SAMPLE

Fresh Serum

GENERAL SYSTEM PARAMETER FULLY AUTO

Mode of reaction Rate


Wavelength 1 570 nm

Wavelength 2 800 nm

Temperature 37oC


Linearity 1000 ng/mL

Blank Reagent Blank

sample volume 3 µL




CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the Ferritin calibrator by the corresponding factor stated in the tablebelow to obtain the Ferritin concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0


Blank calibrator Sample/control

Ferritin R 1 210 µL 210 µL 210 µL



Ferritin R 2 70 µL 70 µL 70 µL

Mix and read absorbance (A1) immediately and after 2 minutes (A2) at 570 nm and800 nm.

CALCULATION




Measuring Range:- 10 –1000 ng/mL.

If the concentration is greater than 1000 ng/mL , dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.

Prozone Effect:- >30000 ng/mL

Precision in CV%:-

Low High

Intra - Run 10 7

Inter - Run 12 10

Accuracy in ng/mL

control Assigned value Measured value|

level 1 31.2(24.9-37.4) 32.6

level 2 193(154-231) 182.2

level 3 331(264-397) 303.67

INTERFERENCE

No interference for



Bilirubin 30 mg/dL

Rheumatoid factor 560 IU/mL

BIBLIOGRAPHY

1. Cook, J.D., Lipschitz,D.A., Laughton, M.B.B., Miles, E.M. & Finch, C.A: Serumferritin as a measure of iron stores in normal subjects. Am.J.clin.Nutr. 27: 680,1974.

2. Walters,G.O.,Miller, F.M & Wormwood, M.: Serum ferritin concentration onand iron stores in normal subjects. J.Clin.Pathol. 26: 770-, 1973.

FERRITIN WITH CALIBRATOR 1 x 30/1 x 10/1 mL

11814002

63

INTENDED USE

This reagent is intended for in vitro quantitative determination of HbA1c in humanblood.


-Ready to use liquid stable reagents


-Direct result (% HbA1c) from analyzer

-No total Hb determination required


HbA1c is a glycated form of haemoglobin formed by the attachment of glucoseresidues in the blood to the hemoglobin molecules. In the diabetic population whereblood glucose levels are abnormally elevated the level of HbA1c also increases. Thelevel of HbA1c is proportional to the level of glucose in the blood and has been widelyaccepted as an indicator of the mean blood glucose concentration in the preceeding6-8 weeks. It is therefore a long-term indicator of diabetic control. For routine useHbA1c levels should be monitored every 3-4 months. However in gestational diabetesand after a change in therapy it may be useful to measure HbA1c more frequently at2-4 week intervals.

PRINCIPLE

This method utilizes the interaction of antigen and antibody to directly determine theHbA1c in whole blood. Total heomoglobin and HbA1c have the same nonspecificabsorption rate to latex particle. When mouse antihuman HbA1c monoclonalantibodies are added (R2), latex HbA1c – mouse antihuman HbA1c antibody complexis formed. Agglutination occurs when goat anti mouse IgG polyclonal antibodyinteracts with the monoclonal antibody. The amount of agglutination is proportionalto the amount of HbA1c absorbed onto the surface of latex particles. The amount ofagglutination is measured as absorbance, which is used to calculate HbA1c % froma calibration curve.

REAGENT COMPOSITION

HbA1c R1 4 x 7.5 mL

Latex0.13%(w/v)

Glycine buffer 20 mmol/L

HbA1c R2A 1 x 9.5 mL


HbA1c R2B 1 x 0.5 mL

Mouse anti-human HbA1c 10 mmol/L

Monoclonal antibody

Goat anti-mouse IgG 0.05 mg/mL

(Polyclonal)

Stabilizers 0.08 mg/dL

HbA1c R3 1 x 60 mL

Haemolysis Reagent

HbA1c DIRECT CALIBRATOR 4 x 0.5 mL

HbA1c 4 level calibrator lyophilized.


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C, protected from light.

MEARUREMENT RANGE

4-16% (NGSP)

NORMAL RANGE



According NGSP :

<6% for non-diabetic

< 7% for glycemic control of person with diabetes



Reagent R2 is prepared by pouring the entire contents of the R2B vial into the R2Avial. Mix gently. This working reagent is stable for 30 days at 2-80C.

Calibrator: Reconstitute with 0.5 mL distilled water and it will be stable up to30 days at 2-80C. Do not freeze.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent and standard bottles immediately after use.


SAMPLE

Whole blood, collected with EDTA

To determine HbA1c a heamolysate must be prepared for each sample

1. Dispense 1mL hemolysis reagent into a tube.2. Add 20 µLof well-mixed whole blood and mix.3. Allow to stand for 5 minutes or until complete lysis is evident.Follow the same procedure with calibrators and controls.

GENERAL SYSTEM PARAMETER FullyAuto



Wavelength 600 nm

Temperature 370C

Calibrator Concentration As on vial label

Linearity 16%

Blank Reagent blank

Sample volume 6 µL






HbA1C R 1 210 µL 210 µL 210 µL

Hemolysate(calib) - 6 µL -

Hemolysate(sample/control) - - 6 µL

Mix & incubate for 5 min at 370C.

HbA1C R 2 75 µL 75 µL 75 µL

Mix and incubate for 5 min at 37oC and read absorbance(A) at 600 nm.

CALCULATION

Calibration curve

Calculate the Abs of calibrators = Abs calibrator – Abs Blank. Plot the D Abs of eachcalibrator versus assigned concentration (HbA1c %) on a linear graph paper. HbA1cresults according to NGSP for the samples and controls are determined using theprepared calibration curve.

Calculate

Abs of sample ie abs Sample - abs Blank .

HbA1c % in the sample is calculated by interpolation of Abs of sample on thecalibration curve. For calculation of results according to IFCC, use IFCC calibratorvalues (see calibrator insert), or use following equation.

NGSP = (0.915 x IFCC) + 2.15

Accuracy in %


level 1 4.8 (3.8-5.9) 4.6

level 2 10.2(8.1-12.3) 10.1

INTERFERENCES

No interference up to :

Ascorbic acid 50 mg/dL

Bilirubin 50 mg/dL

Triglycerides 2000 mg/dL

Carbamylated Hb 7.5 mmol/L

Acetylated Hb 5.0 mmol/L

It has been reported that results may be inconsistent in patients who have thefollowing conditions: opiate addiction, lead poisoning, alcoholism, and ingestion oflarge doses of aspirin. Elevated HbF levels may lead to under estimation of HbA1c.

BIBLIOGRAPHY

1. Nathan, D.M., Clin, Chem. 29, pp.466-469 (1983)2. Engbeak, F., et al. Clin chem.35 pp. 93-97 (1989)3. American Diabetes Association : Clinical practice recommendations (position

statement). Diabetes care 24 (suppl.1) S33-S55, (2001).4. Tietz, N.W. Textbook of Clinical Chemistry, W.B. Saunders Company, p.794

-7795 (1999).

HbA1c DIRECT WITH CALIBRATOR 4 x 7.5/1 x 9.5/1 x 0.5/1x60mL/ 4 x 0.5 mL

11806001

64

INTENDED USE

This reagent is intended for in vitro quantitative determination of IgM antibodies inserum.



-No sample dilution




IgM is important in early response to infections . The measurement of IgM is importantfor typing immuno deficiencies and myelomas. IgM plays an important role in thehumoral defense of the body. Serum levels may be increased in all kind of acuteinfections. Elevated levels in cord serum suggest clinical infection in the new born.

PRINCIPLE

Antibodies to IgM are combined with IgM in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgM in the serum. The light scattering is measuredat 340 nm and 700 nm.

REAGENT COMPOSITION

IgM R1 (Buffer solution) 1 x 30 mL

Tris(hydroxymethyl)aminomethane 100 mmol/L

IgM R2 (Antiserum solution) 1 x 10 mL

Anti-human IgM antiserum

Calibrator 1 x 1 mL



The sealed reagents are stable upto the expiry date mentioned on the label when storedat 2-8 0C. Do not freeze.

REFERENCE RANGE

It is recommended that each laboratory establish its own reference values. Thefollowing values may be used as reference.


PRECAUTION



SAMPLE

Fresh Serum.

GENERAL SYSTEM PARAMETERS SEMI AUTO FULLY AUTO

Mode of reaction Endpoint Endpoint


Wavelength 340/630 nm 340/700 nm





Incubation time(in minutes) 5min+ 5min 5min+ 5min





CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the IgM calibrator by the corresponding factor stated in the tablebelow to obtain the IgM concentration of each


Cal. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



IgM R 1 180 µL 180 µL 180 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance (A1) at 340 nm & 700nm.

IgM R 2 60 µL 60 µL 60 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance(A 2) at 340 nm &700 nm.



IgM R 1 450 µL 450 µL 450 µL

Dil. Calibrator - 5µ L -



IgM R 2 150 µL 150 µL 150 µL

Mix well and incubate for 5 minutes at 37°C. Measure the absorbance(A ) againstreagent blank at 340 nm & 630 nm.

CALCULATION




Measuring Range:- 6 – 500 mg/dL

If the concentration is greater than 500 mg/dl , dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 1.43 1.59 0.64

Inter - Run 3.17 3.65 2.49

Accuracy mg/dL


level 1 63.0(50.4-75.5) 64.1

level 2 140.0(112-168) 143.68

level 3 206(165-247) 209.1

INTERFERENCE

No interference for

Hemoglobin upto 1000mg/dL


Bilirubin upto 20mg /dL

Turbidity upto 5 %

BIBLIOGRAPHY

1. Otani, H. :Medical Technology,14, 965(1986)2. Rinsho Kensa Guide 1992, 269-274(1992), BUNKODO CO. LTD.3. Kanai’s manual of Clinical laboratory Medicine, 30th edition, 868- 873(1993),

KANEHARA & CO.,LTD.

IgM WITH CALIBRATOR 1 x 30/1 x 10/1 mL

11817001


65

INTENDED USE

This reagent is intended for in vitro quantitative determination of IgG antibodies inserum.


-High linearity of 2700 mg/dL




IgG is a predominant serum immunoglobulin . The measurement of IgG is importantfor typing immunodeficiencies and myelomas. Increased levels are found in chronicinfections and chronic inflammation. IgG is the only immunoglobulin which crossesthe placenta and is therefore of special importance in infants defense againstinfection.

PRINCIPLE

Antibodies to IgG are combined with IgG in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgG in the serum. The light scattering ismeasured by reading turbidity at 340 nm.

REAGENT COMPOSITION

IgG R1 (Buffer solution) 1 x 15 mL


IgG R2 (Antiserum solution) 1 x 15 mL

Anti-human IgG antiserum

Calibrator 1 x 1 mL



The sealed reagents are stable upto the expiry date mentioned on the label when storedat 2-8 0C.

REFERENCE RANGE

It is recommended that , each laboratory should establish its own reference values.The following value may be used as a reference.

serum : 650 - 1600 mg/dL

PRECAUTION


SAMPLE

Use fresh serum.








Blank Reagent Bank Reagent Bank

Incubation time 5min + 5min 5min + 5min





PROCEDURE


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the IgG calibrator by the corresponding factor stated in the tablebelow to obtain the IgG concentration of each dilution.


Cal(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



IgG R 1 150 µL 150 µL 150 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance (A1) at 340nm.

IgG R 2 150 µL 150 µL 150 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance(A2) at 340nm.



IgG R 1 250 µL 250 µL 250 µL

Dil. Calibrator - 5µL -



IgG R 2 250 µL 250 µL 250 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance (A) at 340 nm.

CALCULATION




If the concentration is greater than 2700 mg/dL , dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 2.5 3.25 4.18

Inter - Run 4.08 1.83

Accuracy in mg/dL


level 1 967(774-1160) 897.3

level 2 1745(1396-2094) 1803.5

level 3 2643(2114-3171) 2595.7

INTERFERENCE

No interference for




BIBLIOGRAPHY

1. Otani,H. :Medical Technology,14, 965(1986)2. Rinsho Kensa Guide 1992, 269-274(1992), BUNKODO CO., LTD3. Kanai’s manual of Clinical laboratory Medicine, 30th edition, 868-873(1993),

KANEHARA & CO.,LTD.

IgG WITH CALIBRATOR1 x 15/1 x 15/1 mL

11816001


66

INTENDED USE

This reagent is intended for in vitro quantitative determination of IgA antibodies, inserum.



-No sample dilution




The measurement of IgA is important for typing immunodeficiencies and myelomas.Further more it plays a role in acute and chronic infections as first line of defence.Increased levels may be found in acute infectious hepatitis, chronic aggressivehepatitis, cryptogenic cirrhosis, active alcoholic cirrhosis, chronic infections,rheumatoid arthritis and mixed connective tissue diseases etc.

PRINCIPLE

Antibodies to IgA are combined with IgA in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgA in the serum. The light scattering ismeasured by reading turbidity at 700 nm.

REAGENT COMPOSITION

IgA R1(Buffer solution) 1 x 30 mL


IgA R2(Antiserum solution) 1 x 10 mL

Anti-human IgA antiserum

Calibrator 1 x 1 mL




REFERENCE RANGE

It is recommended that , each laboratory should establish its own reference values.The following values may be used as reference.

Serum : 110 - 410 mg/dL

PRECAUTION


SAMPLE

Use fresh Serum

ENERAL SYSTEM PARAMETERS SEMI AUTO FULLY AUTO









Sample volume 5µL 2 µL




CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the IgA calibrator by the corresponding factor stated in the tablebelow to obtain the IgA concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



Ig A R 1 180 µL 180 µL 180 µL




Ig A R 2 60 µL 60 µL 60 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance(A2) at 700 nm.



Ig A R 1 450 µL 450 µL 450 µL




Ig A R 2 150 µL 150 µL 150 µL

Mix well and incubate for 5 minutes at 37°C. Measure the absorbance against thereagent blank at 630 nm.

CALCULATION







Precision in CV%:-

Low Medium High

Intra - Run 0.84 0.94 1.20

Inter - Run 1.70 1.41 1.07

Accuracy in mg/dL


level 1 115(91.7-138) 111.45

level 2 241(192-289) 240.16

level 3 347(277-416) 351.23

INTERFERENCE

No interference for



Bilirubin 20mg /dL

BIBLIOGRAPHY

1. K.Bergstorm, et al.: Scand. J. Clin. Lab. Invest., 637(1980)2. Rinsho Kensa Guide 1992, 269-274(1992), BUNKODO CO. LTD.3. Kanai’s manual of Clinical laboratory Medicine, 30th edition, 868-873(1993),

KANEHARA & CO.,LTD

IgA WITH CALIBRATOR 1 x 30/1 x 10/1 mL

11815001


Calibration curve Spline Spline

67

INTENDED USE

This reagent is intended for in vitro quantitative determination of IgE in human serumand plasma samples.


-Linearity upto 1000 IU/mL


-No need to dilute samples



IgE is an immunoglobulin with a molecular weight of approximately 190 KD. It isnormally present in the blood in trace amounts. IgE, like all immunoglobulins, isproduced by plasma cells in response to antigenic stimuli. However abnormal IgElevels often results in the development of clinically important Type 1 allergicreactions such as asthma, hay fever, dermatitis and food allergies. Elevated levelsare also seen in cases of parasitic infections, pulmonary aspergillosis, Wiskott-Aldrich Syndrome, hepatitis and myeloma.

The measurement of IgE in human serum is considered to be useful in the diagnosis,treatment, assessment of disease progression, or post-operative prognosis for suchconditions.

PRINCIPLE

When an antigen-antibody reaction occurs between IgE and anti-IgE antibody whichhas been coated on latex particles, agglutination results. This agglutination isdetected as an absorbance change, with the magnitude of the change beingproportional to the quantity of IgE in the sample. The actual concentration is thendetermined by interpolation from a calibration curve prepared from calibrators ofknown concentration.

REAGENT COMPOSITION

IgE R1 1 x 32 mL


IgE R2 1 x 8 mL

Latex suspension

0.125 % w/v suspension of latex

particles sensitized with anti IgE

antibodies (mouse)

Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C. DO NOT FREEZE

REFERENCE RANGE



Less than 1 year old 1.35 - 19.5 IU/mL

1 -3 yrs 5.24 - 30.0 IU/mL

4 -6 yrs 5.20 - 112.0 IU/mL

6 - 9 yrs 13.12 - 142.0 IU/mL

10 - 12 yrs 11.2 - 172.0 IU/mL

13 - 18 yrs 25.0 - 126.00 IU/mL

> 19 yrs 28.00 - 140.0 IU/mL

PRECAUTION


SAMPLE

Use Serum / Plasma

GENERAL SYSTEM PARAMETERS SEMI AUTO FULLY AUTO

Mode of reaction Fixed time Fixed time


Wavelength 578nm 570nm/ 800nm





Delay 5 secs -

Delta 120 secs -

sample volume 10 µL 3 µL




CALIBRATION


Prepare the following calibrator dilution using normal saline as diluent. Multiply theconcentration of the IgE calibrator by the corresponding factors stated in the tablebelow to obtain the IgE concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



IgE R 1 200 µL 200 µL 200 µL



IgE R2 50 µL 50 µL 50 µL

Mix and read absorbance (A1)immediately and after 2 minutes (A2) at 570 nm and800 nm.



Ig E R 1 400 µL 400 µL 400 µL



Ig E R2 100 µL 100 µL 100 µL

Mix and measure the absorbance(A1) immediately and after 2 minutes (A2) againstthe reagent blank at 578 nm.

CALCULATION


Calculate the abs and plot a standard curve & read the concentration of controls &samples.


Measuring Range:- 25 –1000 IU/mL.

If the concentration is greater than 1000 IU/mL , dilute the sample with normalsaline and repeat the assay .Multiply the result with dilution factor.

Prozone Effect:- >50000IU/mL

Precision in CV%:-

Low Medium High

Intra - Run 1.18 0.36 0.82

Inter - Run 4.56 1.88 1.21

Accuracy in IU/mL


Control level 1 46.9(37.5-56.3) 44.8

Control level 2 92.5(74-111) 89.8

Control level 3 138(111-166) 134.85

INTERFERENCE

No interference for




BIBLIOGRAPHY

1. Human neutrophils synthesize IL-8 in an IgE-mediated activation Monteseirin, J.et al. J Leukoc Biol. 76: 692-700, 2004

2. Myeloperoxidase release after allergen-specific conjunctival challenge Monteseirin, J. et al. J Asthma. 41: 639-643, 2004

3. IgE-dependent release of myeloperoxidase by neutrophils from allergic patientsMonteseirin, J. et al. Clin Exp Allergy. 31 (6): 889-891, Jun 2001

IgE WITH CALIBRATOR 1 x 32/1 x 8/1 mL

11818002


68

INTENDED USE

This reagent is intended for in vitro quantitative determination of Lipoprotein (a) inserum.

-Multipoint calibration with fixed time mode

-Linearity up to 80 mg/dL


Lp(a) is a low density lipoprotein like particle containing apoliprotein B-100disulphide-linked to one large glycoprotein called apoliprotein(a). Many investigatorshave confirmed that a high lipoprotein(a) concentration represents an indicator of riskfor cardio vascular diseases, especially when, the serum LDL-cholesterol or apo B areelevated. The quantification of Lp (a) in serum or plasma is important for identificationof individuals at risk for developing artherosclerosis.

PRINCIPLE

Latex particles coated with anti-human Lp(a) are agglutinated when mixed with samplescontaining Lp(a). The agglutination causes an absorbance change dependent upon theLp(a) concentration of the patient sample, that can be interpolated in a calibrationcurve prepared with different calibrators of different Lp(a) contents.

REAGENT COMPOSITION

LIPOPROTEIN (a) R1 1 x 20 mL

Buffer solution (pH 8.3)


Lipoprotein (a) latex

LIPOPROTEIN (a) CALIBRATOR 1 x 1 mL

Calibrator concentration as on the vial label


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C. DO NOT FREEZE

NORMAL RANGE



Serum up to 30 mg/dL


Reagent 1 & Reagent 2 are ready to use, should be gently mixed before use.

Calibrator:Reconstitute the calibrator with 1 mL of distilled water & stable for 7 daysat 2-80C.

PRECAUTION


Avoid direct exposure of to light.

SAMPLE

Fresh Serum sample (free of haemolysis)


Mode of Reaction Fixed time with multicalibrator



Temperature 370C

No of calibrators 5

Calibrator Concentration as on the vial label x Dilution factor

Linearity 80 mg/dL

Blank DI water

Delay time 5 sec

Interval 240 sec





CALIBRATION

Calibration curve : Prepare dilutions of the Lp(a) calibrator using 9 g/L saline as diluent:

Dilution 1 2 3 4 5

Cali. (µL) - 25 50 75 100

Saline (µL) 100 75 50 25 -

Dil. factor 0 0.25 0.5 0.75 1.0

Multiply the Lp(a) calibrator concentration by the corresponding dilution factorindicated in the table to obtain the Lp(a) concentration of the different (diluted)calibrators.


Calibrator Sample/Control

R 1 Buffer 800 µL 800 µL

R 2 Buffer 200 µL 200 µL

Calibrator 15 µL -

Sample - 15 µL

Mix and read the absorbance against blank after 10 seconds (A1) and after 4 minutes(A2) of the latex addition.

CALCULATION

Calculate the absorbance differences (A2-A1) of each diluted Lp(a) calibrator and plotthe values against the Lp (a) concentration in a calibration curve. Lp (a) concentrationin the sample is calculated by interpolation of (A2-A1) value on the calibration curve.


1. Measurement Range: 12-80 mg/dLIf the concentration is greater than linearity (80 mg/dL), dilute the sample andrepeat the assay. Multiply the result with dilution factor.

2. Prozone effect: No prozone effect was detected up to 225 mg/dL.

INTERFERENCES

Bilirubin : up to 427 mmol/L no interference

Hemoglobin : up to 10 g/L no interference

Lipids : up to 5 g/L no interference

BIBLIOGRAPHY

1. Gaubalz, J. W. et al. J.Biol Chem 1983; 258 45832 -45892. Berg, K. A. Acta Pathol Microbiol Scand 1963:59:369-3823. Scanu, A. M. et al. J.Clin invest 1990;85: 1709 -1715

LIPOPROTEIN (a) WITH CALIBRATOR 1 x 20/1 x 4/1 mL

11804001

69

INTENDED USE

This reagent is intended for in vitro quantitative determination of microalbumin inurine.

-Turbilatex method with high sensitivity & specificity


-Single point calibration


The significant increase of albumin concentration in the urine has been used someyears as an indicative value of incipient nephropathy and cardiovascular disease indiabetic patients. Microalbuminuria has also been associated with hypertension andrisk of cardiovascular disease in non-diabetic patients. Microalbuminuria occurs inresponse to acute inflammatory conditions such as ischemia, trauma and thermalinjury, surgery, pancreatitis and inflammatory bowel disease. In many of theseconditions, albumin excretion increase within minutes or hours of the initiatingstimulus. The degree of microalbuminuria is proportional to the severity of theinflammatory process.

PRINCIPLE

Latex particles coated with anti human albumin are agglutinated when mixed withsample containing albumin. The agglutination causes an absorbance change,dependent upon albumin concentration of the patient sample, that can be quantifiedby comparison with a calibrator of known microalbumin concentration

REAGENT COMPOSITION

MICROALBUMIN R1 1 x 45 mL

Diluent)

Glycine buffer (pH 10.0) 100 mmol/L

Sodium azide

MICROALBUMIN R2 1 x 5 mL

Latex

Suspension of latex particles

Coated with anti human

Albumin (pH 8.2)


MICRO ALBUMIN CALIBRATOR 1 x 1 mL

Microalbumin calibrator concentration is stated on the vial label.

Components of human serum have been tested and found to be negative for thepresence of HBs Ag, HCV and of antibody to HIV (1/2). However handle cautiouslyas potentially infectious.


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C. DO NOT FREEZE.

NORMAL RANGE



Urine : up to 15 mg/L


Reagent 1 and reagent 2 are ready to use.

Calibrator : Ready to use. The calibrator is stable until the expiry date on the label whenstored tightly closed at 2-80C.

PRECAUTION



SAMPLE

Fresh urine.

MICROALBUMIN TURBILATEX WITH CALIBRATOR1 x 45/1 x 5/1 mL

11805001


Mode of reaction Fixed time Rate


Wavelength 546 nm 546nm


Calibrator concentration As on vial label As on vial label

Linearity 150mg/L 150mg/L


Delay 2sec --

Interval 120 sec --

Sample volume 7µL 3 µL

Reagent 1 volume 900 µL 270µL

Reagent 2 volume 100 µL 30µL




MAlb R1 270 µL 270 µL 270 µL

Calibrator 3 µL -

Sample /control - - 3 µL


MAlb R2 30 µL 30 µL 30 µL




M Alb R1 900 µL 900 µL

Calibrator 7 µL -

Sample - 7 µL

Malb R2 100 µL 100 µL


CALCULATION

(A2-A1) sample

Malb Conc. in mg/L = ------------------------- x Calibrator Conc.

(A2-A1) calibrator


Measuring Range:- 2 - 150 mg/L.

If the concentration is greater than 150 mg/L. dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 2.25 1.48 1.93

Inter - Run 2.28 2.06 2.55

Accuracy in mg/L


level 1 19.6(15.7-23.5) 21.2

level 2 67.2(53.7- 80.6) 69.8

INTERFERENCE



Creatinine 3 g/L

Bilirubin 10 mg/dL

BIBLIOGRAPHY

1. Feldt –Rasmussen, B. et al J Diab Comp 1994; 8; 137 1452. Panuyiotou, B. N. Journal international Medical Research 1994 ; 22;181,

181-2013. Bar, J. et al Diabetic Medicine 1995; 12;649-6564. Gilbert ,R. E. et al Diabetic Medicine 1994; 11;636 - 6455. Medcalf, E. A. et al. Clin chem. 1990; 36/3; 446-4496. Young, D. S. Eeffects of drugs on clinical laboratory test, 4th ed. AACC Press, 1995.


70

INTENDED USE

This reagent is intended for in vitro quantitative determination of microalbumin inhuman urine

-Turbidometric Immunoassay



-No need to dilute samples



Albumin is normally found in the blood. When the kidneys are working properly,albumin will not be present in the urine. However, when the kidneys are damaged,small amounts of albumin leak into the urine. This condition is calledmicroalbuminuria.

Microalbuminuria is most often caused by kidney damage from diabetes. However,many other conditions can lead to kidney damage, such as high blood pressure,heart failure, cirrhosis, or systemic lupus erythematosus (SLE). If early kidneydamage is not treated, larger amounts of albumin and protein may leak into theurine. This condition is called macroalbuminuria or proteinuria, this can lead tochronic kidney disease.

PRINCIPLE

The reagents containing polyclonal goat antihuman microalbumin when mixed withthe urine sample containing microalbumin cause changes in absorbance, due tothe development of turbidity, which is directly proportional to the concentrationof microalbumin in the sample.

REAGENT COMPOSITION

Microalbumin R1 2 x 25 mL

Saline (9 g/L)

Accelerator



Phosphate buffered saline

Polyclonal goat anti-human albumin (variable)


Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C. DO NOT FREEZE!

NORMAL RANGE



Urine : 0 -25 mg/L (IFCC)

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent bottles immediately after use. Avoid direct exposurereagent to light.Do not Freeze.

SAMPLE

Use fresh Urine.

GENERAL SYSTEM PARAMETERS FULLY AUTO

Mode of reaction Endpoint


Wavelength 340 nm

Temperature 37o C

Calibrator concentrationAs on vial label x Dilution factor

Linearity 395 mg/L

Blank Reagent Blank

Incubation time 5min + 5 min

Sample volume 3 µL




CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the microalbumin calibrator by the corresponding factor stated inthe table below to obtain the microalbumin concentration of each dilution.


Calib.(µL) - 10 10 25 50 100

Saline(µL) 100 150 70 75 50 -

Dil. Factor(µL) 0 0.0625 0.125 0.25 0.5 1.0


Blank Calibrator Sample /control

Microalbumin R 1 200 µL 200 µL 200 µL



Mix and incubate for 5 minutes at 37°C. Read the absorbance (A 1) at 340 nm.

Microalbumin R2 40 µL 40 µL 40µL


CALCULATION




Measuring Range:- 4 - 395 mg/L

If the concentration is greater than linearity (395 mg/L), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 2.28 1.8 3.04

Inter - Run 2.93 0.66 0.53

Accuracy in mg/L


level 1 19.6(15.7-23.5) 24.0

level 2 67.2(53.7- 80.6) 62

INTERFERENCE

No interference for

Hemoglobin upto 1000 mg/dl


BIBLIOGRAPHY

1. Mount, J. J. Clin. Pathology, 22, 12 (1986)2. Schmidtz, A. et al., diabetic Medicine, 5 , 126 (1988)

MICROALBUMIN IT WITH CALIBRATOR 2 x 25 /2 x 5/1 mL

11824001

ADL/V.02/Februay 2013

71

INTENDED USE

This reagent is intended for in vitro quantitative determination of Prealbumin inhuman serum





Prealbumin is a serum and cerebrospinal fluid carrier of thyroid hormone thyroxine(T4) and retinol. So the more accurate name for prealbumin is transthyretin .

Prealbumin is formed in the liver, it is a measure of hepatocyte function. Decreasedand increased levels of serum prealbumin are associated with liver disease and areaffected by the existence and degree of liver diseases.

The half life of prealbumin is approximately 2 days, making prealbumin a more timelyand sensitive indicator of protein status.

PRINCIPLE

Antibodies to prealbumin are combined with prealbumin in the patient’s serum,forming immune complexes.The immune complexes cause an increase in the lightscattering with the concentration of prealbumin in the serum. The light scattering ismeasured at 340 nm and 700 nm.

REAGENT COMPOSITION

Prealbumin R1(Buffer Solution) 1 x 30 mL

Tris (hydroxymethyl) aminomethane 100 mmol/L

Prealbumin R2 (Anti serum solution) 1 x 3 mL

Anti- human prealbumin anti serum(Variable)

Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C. DO NOT FREEZE.

NORMAL RANGE





PRECAUTION


SAMPLE















CALIBRATION


Dilute the high concentrated calibrator to 1/10 using normal saline and use thisdiluted calibrator for the preparation of calibration curve. Prepare the followingcalibrator dilutions using NaCl as diluent. Multiply the concentration of thePrealbumin calibrator by the corresponding factor stated in the table below to obtainthe Prealbumin concentration of each dilution.


Cali. 1/10 dil(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0



Prealbumin R 1 300 µL 300 µL 300 µL


Dil. Sample/control - - 15 µL



Mix and incubate for 5 minutes at 37°C. Measure the absorbance (A 2) at 340 nm.

ALTERNATIVE PROCEDURE FOR SEMIAUTOANALYZER:







Mix and incubate for 5 minutes at 37°C. Measure the absorbance at 340 nm.

CALCULATION




If the concentration is greater than 80 mg/dL , dilute the diluted(1/10)sample withnormal saline and repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 5.30 3.04 3.96

Inter - Run 6.22 4.71 4.1

Accuracy in mg/dL


level 1 15.0(12.0-17.9 15.86

level 2 24.7(19.7-29.6) 26.1

level 3 32.1(25.7-38.6) 33.2

INTERFERENCE




Bilirubin 20 mg/dL

BIBLIOGRAPHY

1. Japanese Journal of Clinical Medicine,53(enlarged) ,186 – 188 (1995)2. Journal of Clinical Experimental Medicine, 149 (5), 277 – 279 (1989)

PREALBUMIN WITH CALIBRATOR 1 x 30/1 x 3/1 mL

11823002

72

INTENDED USE

This reagent is intended for in vitro quantitative determination of Rheumatoid factorin serum.

-Linear up to 100 IU/mL



Rheumatoid factor are a group of antibodies directed to determinants in the Fc portionof the immunoglobulin G molecule. Although rheumatoid factors are found in anumber of rheumatoid disorders, such as Systemic Lupus Erythematosus(SLE) andSjogren’s syndrome as well as in non rheumatic conditions. Its central role in clinicslies in its utility as an aid to the diagnosis of rheumatoid arthritis (RA).

PRINCIPLE

The RF agglutination assay is a quantitative turbidimetric assay for measurement ofRF in human serum.

Latex particles coated with human gamma-globulin are agglutinated when mixed withsamples containing RF. The agglutination causes an absorbance change, dependentupon the RF concentration of the patient sample, that can be quantified bycomparison with a calibrator of known RF concentration.

REAGENT COMPOSITION

RF TURBILATEX R1 1 x 45 mL

Diluent

Tris buffer 20mmol/L


RF TURBILATEX R2 1 x 5 mL

RFLatex:


human gamma-globulin. (pH 10.0)


RF CALIBRATOR 1 X 2 mL


STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-8oC. DO NOT FREEZE.

NORMAL RANGE


Serum : up to 20 IU/mL


Reagent R1 and Reagent R2 are ready to use.

RF calibrator: Reconstitute with 2mL distilled water. Stable for 30 days at 2-80C.

Dilution of calibrator for calibration curve.

Prepare the following calibrator dilutions with normal saline as diluent. Multiply theconcentration of the RF calibrator by the corresponding factor stated in the table toobtain RF concentration of each dilution.


Cali. (µL) - 10 30 50 75 100

Saline (µL) 100 90 70 50 25 -

Dil. factor 0 0.1 0.3 0.5 0.75 1.0

For single point calibration (linear up to 100IU/mL): Prepare the RF calibrator dilutionas follows.

30µLRF calibrator + 70µL normal saline

Multiply the RF calibrator concentration by 0.3 to obtain the concentration of thediluted calibrator.

RF TURBILATEX WITH CALIBRATOR 1 X 45 mL/1 X 5 mL/1 X 2 mL11803001

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure of reagentsto light.

SAMPLE



Mode of reaction End point



Temperature 37oC

Calibrator concentration As on vial label x 0.3

Linearity 100 IU/mL

Blank Reagent blank

Incubation time 2 minnutes

Sample volume 5 µL

Reagent1 volume 900 µL

Reagent R2 volume 100 µL




R1 reagent 900 µL 900 µL 900 µL



Reagent R2 100 µL 100 µL 100 µL

Mix and incubate exactly 2 minutes at 37oC. Measure the absorbance of calibratorand sample against reagent blank.

CALCULATION


RF concentration in IU/mL =(Abs of sample /Abs of calibrator) x dil. calib con.


Calculate the difference in absorbance of each diluted calibrator from reagent blankand plot the values obtained against the RF concentration of each dilution on thecalibration curve.

RF concentration in the sample is calculated by interpolation (abs sample-abs blank)in the calibration curve.


Measuring Range

Multipoint calibration : 20-160 IU/mL

Single point : up to 100 IU/mL.


Prozone effect: No prozone effect was detected up to 800 IU/mL.

INTERFERENCES

No interference for

Bilirubin --- up to 20mg/dL

Hemoglobin --- up to 10 g/L

Lipids --- up to 10 g/L

BIBLIOGRAPHY

Frederick Wolfe et al; Arthritis and Rheumatism 1991: 34: 951-960.


73

INTENDED USE

This reagent is intended for in vitro quantitative determination of Rheumatoid factorin Serum.


-Linear upto 135 IU/mL




Rheumatoid Factor (RF) is an auto antibody against human IgG commonly seen in seraof patients with rheumatoid arthritis. The measurement of RF value is useful inevaluating the diagnosis, effects of therapy and prognosis of RA, systemic lupuserythematosus, Chronic hepatopathy etc.

PRINCIPLE

When a sample containing rheumatoid factor is added to denatured human IgG whichhas been sensitized to latex particles, antigen-antibody reaction occurs leading toagglutination. This agglutination leads to an absorbance change which is measured at(550 to 660nm). The change in absorbance is proportional to agglutination and theactual concentration is determined by interpolation from a calibration curve preparedfrom known value calibrators.

REAGENT COMPOSITION

RF R 1 1 x 24 mL, 2 x 24 mL

Glycine Buffer Solution

RF R2 1 x 8 mL, 2 x 8 mL

Latex suspension coated with denatured human IgG

CALIBRATOR 1 x 1 mL, 1 x2 mL

RF calibrator concentration as on vial label.


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C. DO NOT FREEZE.

NORMAL RANGE



Serum up to 18 IU/mL

PREPARATION OF REAGENT

Reagent 1 and Reagent 2 are ready to use

RF Calibrator : Ready to use Liquid stable

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure of reagentto light & heat.


SAMPLE

Fresh serum (Do not use haemolized or lipemic serum)


Mode of Reaction Fixed Time End point




No.of calib. 6 6




Delay 5 sec --

Interval 120 sec --





CALIBRATION


Prepare the following calibrator dilutions using normal saline as diluent. Multiplythe concentration of the RF calibrator by the corresponding factor stated in the tablebelow to obtain the RF concentration of each dilution.


Cali. (µL) - 10 20 50 75 100

Saline (µL) 100 70 60 50 25 -

Dil. factor 0 0.125 0.25 0.5 0.75 1.0


Blank Calibrator Sample /control

RF R1 210 µL 210 µL 210 µL




RF R2 70 µL 70 µL 70 µL




RF R1 450 µL 450 µL


Sample /control - 10 µL

RF R2 150 µL 150 µL


CALCULATION


Calculate the abs, plot a standard curve & read the concentration of controls &samples,by interpolation of curve.


Measuring Range:- 10 - 135 IU/mL.

If the concentration is greater than 135 IU/mL, dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.

Prozone Effect:- >770 IU/mL

Precision in CV%:-

Low High

Intra - Run 5.0 1.0

Inter - Run 8.0 5.0

Accuracy in IU/mL


level 1 23.8(19-28.5) 20.1

level 2 28.4(22.7-34) 30.81

level 3 37.9(30.3-45.5) 41.96

IINTERFERENCE



Intrafat 5000 mg/dL

Bilirubin 20 mg/dL

BIBLIOGRAPHY

Frederick Wolfe et al-Arthritis and Rheumatism 1991 : 34:951-960

RF LEIT WITH CALIBRATOR 1x 24 /1 x 8 / 1 mL, 2x 24 /2 x 8 / 2 mL

11809003, 11809004

74

INTENDED USE

This reagent is intended for in vitro quantitative determination of Transferrin inhuman serum






Transferrin(TF) is synthesised in Liver. The main role of Transferrin is to deliver ironfrom absorption centres to all tissues. Increased serum Transferrin is associated withiron deficiency anemia, malignant tumor, polycythemia rubra, aplastic anemia,hemolytic anemia and hepatitis. Absence of Transferrin in the body creates a raregenetic disorder known as atransferrinemia; a condition characterized by anemia andhemosiderosis in the heart and liver that leads to many complications including heartfailure.

PRINCIPLE

Antibodies to human TF are combined with transferrin in the patients serum, formingimmune complexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of TF in the serum. The light scattering is measured340 nm.

REAGENT COMPOSITION

Transferrin R1(Buffer solution) 1 x 30 mL


Transferrin R2(Anti serum solution) 1 x 3 mL

Anti- human TF anti serum (Variable)

Calibrator 1 x 1 mL



The reagents are stable until expiry date when kept at 2-80 C .

NORMAL RANGE



Male : 190 - 300 mg/dL


PRECAUTION


SAMPLE

Use fresh serum. Dilute serum/control to 1/10 with saline.









Incubation time 5min + 5min -

Sample volume 15 µL 8.5 µL




CALIBRATION


Dilute the high concentrated calibrator 1/10 with normal saline and use this dilutedcalibrator for the preparation of calibration curve.

Prepare the following calibrator dilutions using normal saline as diluent. Multiply theconcentration of the Transferrin calibrator by the corresponding factors stated in thetable below to obtain the Transferrin concentration of each dilution.


1/10dilcali.(µL)| - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1



Transferrin R1 300 µL 300 µL 300 µL

Dil. Calibrator - 8.5 µL -

Dil. Sample/control - - 8.5 µL



Mix and incubate for 5 minutes at 37°C. Measure the absorbance (A 2) at 340 nm.

ALTERNATIVE PROCEDURE FOR SEMIAUTOANALYZER:






Transferrin R 2 50 µL 50 µL 50 µL

Mix and incubate for 5 minutes at 37°C. Measure the absorbance (A) at

340 nm against reagent blank.

CALCULATION



Measuring Range:- 40 – 540 mg/dL.

If the concentration is greater than 540 mg/dL , dilute the diluted(1/10)sample withnormal saline and repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 4.78 1.30 0.78

Inter - Run 4.64 2.46 0.9

Accuracy in mg/dL


level 1 151(121-181) 160.48

level 2 250(200-300) 254.54

level 3 334 (267 - 400) 351.3


No interference for



Lipemia upto5 % of intralipid

BIBLIOGRAPHY

1. K. Bergstorm, et al.: Scand. J.Clin Lab Invest., 40, 637- 640 (1980)2. H. Maikus, et al.: Clinica Chemica Acta, 88, 523- 530(1978)

TRANSFERRIN WITH CALIBRATOR 1 x 30/1 x3/1 mL

11821002

75

INTENDED USE: This reagent is intended for in vitro qualitative & semi quantitativedetermination of Anti Streptolysin O (ASO) in serum.

-No prozone effect detected up to 1500 IU/mL

-Rapid procedures; test time only 2 minutes

-Excellent clarity; clear agglutination


Streptolysin O is a toxic immunogenic exoenzyme produced by ß-haemolyticStreptococcus group A, C and G. ASO antibodies are useful for the diagnosis ofrheumatoid fever, acute glomerulonephritis and streptococcal infections. Rheumaticfever is an inflammatory disease affecting connective tissue from several parts ofhuman body as skin, heart, joints etc. Acute glomerulonephritis is a renal infectionthat affects mainly glomerulus.

PRINCIPLE

ASO latex kit is a rapid agglutination procedure for the direct detection and semi-quantitation (on slide) of antistreptolysin-O (ASO). The antigen, latex particlessuspension coated with Streptolysin-O, agglutinates in the presence of specificantibodies present in the sera of patients with streptolysin O.

REAGENTS & MATERIALS PROVIDED

ASO LATEX 1 x 2.5 mL / 1 x 5 mL

Suspension of polystyrene particles coated with Streptolysin-O.

ASO POSITIVE CONTROL 1 x 0.5 mL

Human pooled serum

ASO NEGATIVE CONTROL 1 x 0.5 mL

Human pooled serum

PHYSIOLOGICAL SALINE BUFFER CONCENTRATE 1 x 5 mL

Dilute 1:20 (v/v) with distilled water

ACCESSORIES: for 50T for 100 T

1. Reaction slide 1 1

2. Serum droppers 50 100

3. Applicator sticks 50 100

4. Rubber teat 1 1

PRECAUTION

Reagent components of human origin have been tested and found to be negativefor the presence of antibody to HIV (1/2) as well as for HBsAg and HCV antibody.However, handle cautiously as potentially infectious.

The reagent and controls contain less than 0.1% sodium azide.


When stored at 2-80C and protected from light, the reagent and the controls arestable until the expiry date stated on the label. DO NOT FREEZE

Preparation of physiological saline buffer: Prepare physiological saline buffer byadding 95mL of distilled water to 5 mL of physiological saline buffer concentrate(provided). It is stable up to expiry date, when stored at room temperature.

ANALYTICAL SENSITIVITY

The ASO latex reagent sensitivity has been adjusted to detect a minimum of

200 (+/- 50) IU/mL ASO antibodies in the undiluted samples.

SAMPLE


QUALITATIVE TEST

Allow all reagents as well as the sample to reach room temperature. Mix well beforeuse.

1. Place 1 drop of serum sample on the slide using a disposable serum dropper.2. Add one drop of ASO-latex reagent to the above drop and mix with disposable

applicator stick.3. Rock the slide gently to and fro for 2 minutes and immediately examine under

a good light source for agglutination, do not examine beyond 2 minutes.4. For positive & negative controls follow the same procedure as mentioned above

by taking control serum from respective vials.RESULT AND INTERPRETATION

Positive result:

The presence of agglutination indicate concentration of ASO in the sample equalor greater than 200 IU/mL (above normal).

Negative result:

The lack of agglutination indicates ASO level lower than 200 IU/mL in the sample,(within the normal range).

ASO50 T, 100 T

12201002, 12201003

SEMI – QUANTITATIVE TEST

In the cases in which it is desired to find out the titre of a positive sample, it is possibleby the serial dilution methodology.

1. Place 50 µL diluted physiological saline Buffer onto each of five circles of the slide2. Using a 50 µL micro pipette add 50 µL of the serum sample to the drop of saline

buffer in 1st circle.3. Using the same micro pipette, mix the sample with saline by aspirating back &

forth several times. Aspirate 50 µL from 1st circle and transfer to 2nd circle.Repeat the same operation up to 5th circle. Aspirate 50 µL from 5th circle anddiscard. Dilutions obtained as 1/2, 1/4 1/8, 1/16, 1/32

4. Then add 1 drop of ASO latex to the above circles. Mix and rock the slide gently toand fro for 2 minutes; observe the agglutination under good source of light.

CALCULATION

ASO Conc. (IU/mL) = sensitivity x Titre (Highest dilution serum showing agglutination)

Where, ASO sensitivity = 200 IU/mL

NOTE:

1. False positive results may be obtained in conditions such as rheumatoid arthritis,scarlet fever, several other streptococcal healthy carriers.

2. Early infections and children from 6 months to 2 years may cause false negativeresults.

3. A single ASO determination does not produce much information about theactual state of the disease. Titrations at bi weekly intervals during 4 or 6 weeksare advisable to follow the disease evolution.


1. Diagnostic sensitivity : 98%2. Diagnostic specificity : 97%3. Prozone effect: No prozone effect detected up to 1500 IU/mL.Interferences

Hemoglobin : 10 g/L

Bilirubin : 20 mg/dL

Rheumatoid factor : 300 IU/mL

do not interfere. Other substances may interfere4.

BIBLIOGRAPHY

1. CURTIS G. D.W, KRAAK W.A.G., MITCHELL R.G. Comparison of latex andhemolysis test determination of antistreptolysin O (ASO) antibodies. J. Clin.Pathol. 41, 1331 (1988)

2. TADZYNSKY L.A., RYAN M.E. Diagnosis of rheumatoid fever. A guide to criteriaand manifestations. Post grad Med. 79, 295 (1986)

3. D’ANGELO W.A. Rheumatic Diseases. Diagnostic tests and procedures. in: TheLaboratory in Clinical Medicine (Halsted J.A. Ed.) Saunders Company,Philadelphia, chapter 29 (1976)

4. Young D.S., Effects of Drugs on Clinical Laboratory Test 4th ed. AACC Press, 1995.


76

INTENDED USE: This reagent is intended for in vitro quantitative determination ofC-reactive protein (CRP) in serum

-No prozone effect detected up to 1600 mg/L

-Rapid procedure, only 2 minutes test

-Excellent clarity, clear agglutination


CRP is a classic acute phase protein of human serum, synthesized by hepatocytes.Normally it is present only in trace amounts in serum, but it can increase as muchas 1,000 fold in response to injury or infection. The clinical measurement of CRP inserum therefore appears to be a valuable screening test for organic disease and asensitive index of disease activity in inflammatory, infections and ischemic conditions.

PRINCIPLE

CRP latex kit is a rapid agglutination procedure for the direct detection and semi-quantitation (on slide) of C-reactive protein (CRP). The reagent, latex particlessuspension coated with specific antihuman C-reactive protein antibodies,agglutinates in presence of CRP in patient serum.


CRP LATEX 1 x 2.5 mL / 1 x 5 mL

Suspension of polystyrene particles coated with anti human CRP goat antibodies.

CRP POSITIVE CONTROL 1 x 0.5 mL

Human pooled serum

CRP NEGATIVE CONTROL 1 x 0.5 mL

Human pooled serum


Dilute 1:20 (v/v) with distilled water

ACCESSORIES: For 50 T For 100 T

1. Reaction slide 1 1

2. Plastic droppers 50 100

3. Applicator sticks 50 100

4. Rubber teat 1 1

PRECAUTION




When stored at 2-80C and protected from light, the reagent and the controls arestable until the expiry date stated on the label. DO NOT FREEZE.

Preparation of physiological saline buffer: Prepare physiological saline buffer byadding 95mL of distilled water to 5mL of physiological saline buffer concentrate(provided). It is stable up to expiry date, when stored at room temperature.


The CRP latex sensitivity has been adjusted to detect a minimum of6 (5-10) mg/L in the undiluted samples.

SAMPLE


QUALITATIVE TEST


1. Place 1 drop of serum sample on to the slide using a disposable serum dropper.2. Add one drop of CRP-latex reagent to the above drop and mix well with

disposable applicator stick.3. Rock the slide gently to and fro for 2 minutes and examine immediately under

good light source for agglutination, do not examine beyond 2 minutes.4. For positive & negative controls follow the same procedures as mentioned

above by taking control serum from respective vials.RESULT AND INTERPRETATION

Positive result:

The presence of agglutination indicate concentration of CRP in the sample equalor greater than 6 mg/L (above normal)

Negative result:

The lack of agglutination indicates CRP level lower than 6 mg/L in the sample,(within the normal range)

CRP 50 T, 100 T

12202002, 12202003


In the case in which it is desired to find out the titre of a positive sample, it ispossible by the serial dilution methodology.

1. Place 50 µL diluted saline Buffer onto each of five circles of the slide.2. Using a 50 µL micro pipette add 50 µL serum sample to the drop of saline


forth several times. Aspirate 50 µL from 1st circle and transfer to 2nd circle.Repeat the same operation up to 5th circle. Aspirate 50 µL from 5th circle anddiscard. Dilutions obtained as 1/2, 1/4, 1/8, 1/16, 1/32

4. Then add 1 drop of CRP latex reagent to the above circles. Mix and rock the slidegently to and fro for 2 minutes; observe the agglutination under good source oflight.

CALCULATION

Concentration of CRP in serum can be calculated as follows:

CRP Conc. (mg/L) = sensitivity x titre (highest dilution serum showing agglutination)

Where, CRP sensitivity = 6 mg/L

NOTE:

1. Reaction time is critical, if reaction time exceeds two minutes, drying of reactionmixture may cause false positive results.

2. Freezing the CRP latex reagent will cause spontaneous agglutination.3. Intensity of agglutination is not necessarily indicative of relative CRP

concentration, therefore screening reaction should not be graded.4. A false negative can be attributes to a prozone phenmenon (antigen excess). It

is recommended therefore to check sample with dilution.5. Clinical diagnosis should not be made on findings of a single test result, but

should integrate both clinical and laboratory data.


Diagnostic sensitivity : 95.6%

Diagnostic specificity : 96.2%

Prozone effect : No prozone effect detected up to 1600mg/mL.

INTERFERENCES


Hemoglobin up to 10 g/L

Lipids up to 10 gL

Rheumatoid factor up to 100 IU/mL

Do not interfere. Other substances may interfere.

BIBLIOGRAPHY

1. Wadsworth, C.Wadsworth, E., Efficacy of latex agglutination methods fordetermination of C - reactive protein in Pediatric sera. Clin. Chem. Acta, 138,(1984), 309

2. Ballou S.P, Kushner, I.C. Reactive Protein and acute phase response. ADV Int.Med, 37, (1992), 313.

3. Young D.S., Effects of Drugs on Clinical Laboratory Test 4th ed. AACC Press, 1995.


77

INTENDED USE: This reagent is intended for in vitro qualitative & semiquantitative determination of Rheumatoid factor (RF) in serum.

-Sensitivity 8 IU/mL

-Optimized antigen concentration; No prozone effect up to 800 IU/mL

-Rapid procedure; only 2 minutes test

-Excellent clarity; crystal clear agglutination


Rheumatoid factors are a group of antibodies directed to the determinants in the Fcportion of the immunoglobulin G molecule (IgG). Although rheumatoid factors arefound in a number of rheumatoid disorders, such as systemic lupus erythematosus(SLE) and Sjogrens syndrome as well as in non rheumatic condition, its central rolein clinics lies in its utility as an aid in the diagnosis of rheumatoid arthritis (RA).

PRINCIPLE

RF latex kit is a rapid agglutination procedure for the detection and semi-quantitation (on slide) of Rheumatoid Factors (RF). The antigen, a latex particlessuspension coated with human gamma-globulin, agglutinates in presence ofrheumatoid factors in the patient serum


RF LATEX 1 x 2.5 mL / 1 x 5 mL

Suspension of polystyrene particles coated with human gamma -globulin.

RF POSITIVE CONTROL 1 x 0.5 mL

Human pooled serum

RF NEGATIVE CONTROL 1 x 0.5 mL

Human pooled serum


Dilute 1:20(v/v) with distilled water

ACCESSORIES:For 50 T For 100 T

1. Reaction slide 1 12. Plastic droppers 50 1003. Applicator sticks 50 1004. Rubber teat (blue) 1 1PRECAUTION





Preparation of physiological saline buffer: Prepare saline buffer by adding 95 mLof distilled water to 5 mL of concentrate saline buffer (provided). It is stable up toexpiry date, when stored at room temperature.


The RF sensitivity has been adjusted to detect a minimum of

8.0 IU/mL (6-16) RF in the undiluted samples.

SAMPLE


QUALITATIVE TEST


1. Place 1 drop of serum sample on to the slide with disposable serum dropper.2. Add one drop of RF-latex reagent to the above drop and mix well with

disposable applicator stick.3. Rock the slide gently to and fro for 2 minutes and then examine immediately

under good light source for agglutination, do not examine beyond 2 minutes.4. For positive & negative controls follow the same procedures as mentioned

aboveby taking control serum from respective vials.RESULT AND INTERPRETATION

Positive result:

The presence of agglutination indicated concentration of RF in the sample equalor greater than 8 IU/mL (above normal).

Negative result:

The lack of agglutination indicates RF level lower than 8 IU/mL in thesample, (within the normal range).


In the case in which it is desired to find out the titre of a positive sample, it isfeasible by the serial dilution methodology.

1. Place 50 µL diluted saline buffer onto each of five circles of the slide2. Using a 50 µL micro pipette add 50 µL of the serum sample to the drop of saline


forth several times. Aspirate 50 µL from 1st circle and transfer to 2nd circle.Repeat the same operation up to 5th circle. Aspirate 50 µL from 5th circle anddiscard. Dilutions obtained as ½, ¼, 1/8, 1/16, 1/32

4. Then add 1 drop of RF latex reagent to the above circles, mix and rock the slidegently to and fro for 2 minutes; observe the agglutination under good sourceof light.

CALCULATION

The Rheumatoid Factor (RF) level in serum can be calculated as follows:

RF conc. (mg/dL) = sensitivity x titre (highest dilution of serum showing agglutination)

Where, RF Sensitivity = 8.0 IU/mL


1. Diagnostic sensitivity : 98%

2. Diagnostic specificity : 98.8%

3. Prozone effect : No prozone effect was detected up to 800 IU/mL.

INTERFERENCES


Hemoglobin up to 10g/L

Lipids up to 10g/L

will not interfere. Other substances may interfere.

NOTE:

1. The incidence of false positive results is about 3-5%. Individuals suffering frominfectious mononuclear hepatitis, syphilis as well as elderly people may givepositive results.

2. Diagnosis should not be solely based on the results of latex method, but alsoshould be complemented with a Waaler Rose test along with clinicalexamination.

BIBLIOGRAPHY

1 . Frederick Wolf et al-Arthritis and Rheumatism 1991: 34: 951-9602. Robert W Dorner et al. Clinica Chemica Acta 1987; 167: 1-21

RF 50 T, 100 T

12203002, 12203003


78

INTENDED USE: RPR (Rapid Plasma Reagin) test is a non-treponemal test for theserological diagnosis of Syphilis. Specificity, reactivity and sensitivity are similar tothat of classical VDRL test. It is a reliable, economical and rapid test and hencerecommended as a screening test.


Syphilis is a veneral disease caused by the spirochete microorganism T.pallidum. Asthe organism is difficult to be cultured on artificial media, the diagnosis of syphilisdepends on the correlation of clinical data with the detection of specific antibodyby serological test. Serological screening tests for syphilis using cardiolipin andlecithin antigen are simple to perform and reliable but may give rise to smallproportion of false positive results because the tests use non treponemal antigens.These antigens detect reagin antibodies produced against cardiolipin during aninfection.

PRINCIPLE

RPR reagent is an antigen containing cardiolipin antigen, sensitized on carbon particleswhich detects reagin antibody present in serum of syphilitic individuals. When aspecimen contains antibody is mixed with the reagent, flocculation occurs due tocoagglutination of the carbon particles of the RPR antigen, which appear as blackclumps against the white background of the card. This coagglutination is readmacroscopically. Non-reactive specimens show a black button at the centre.

REAGENTS AND METERIALS PROVIDED

RPR ANTIGEN 1 x 1.1 mL, 1 x 2.6 mL, 4 x 2.6 mL

Antigen, Cardiolipin suspension containing carbon micro particles.

RPR POSITIVE CONTROL 1 x 0.3 mL

Human pooled serum

RPR NEGATIVE CONTROL 1 x 0.3 mL

Human pooled serum

PHYSIOLOGICAL SALINE 1 x 5mL

Dilute 1:20 (v/v) with H2O.

ACCESSORIES: For 50 Tests For 125 Tests For 500 Tests

1.Applicator stick 50 125 500

2. Serum dropper 50 125 500

3. Plastic slides 7 16 63

4. Dropper with needle 1 set 1 set 1 set

( Accessories will be provided in separate pouch)

PRECAUTION





Preparation of saline buffer: Prepare saline buffer by adding 95 mL distilled waterto 5 mL concentrated saline buffer. It is stable up to expiry date when stored atroom temperature.

SAMPLE


QUALITATIVE TEST

Bring the test reagents and samples to room temperature

1. Place 1 drop of serum sample (50 µL or 0.05mL) on to the slide with disposableserum dropper.

2. Place one drop of RPR antigen suspension (18 µL) by antigen dropper3. Mix well and spread the liquid over the entire area of the circle using disposable

applicator stick4. Rock the slide gently to and fro for 8 minutes on a mechanical rotator& observe

immediately under good light source for the appearance of carbon particleclumping.

RESULT AND INTERPRETATION

Medium & large aggregates against white back ground - - Reactive

Small agglutinates around periphery - Weak reactive

No aggregates, button formation at the centre of circle - Non reactive


In the case in which it is desired to find out the titre of a positive sample, it ispossible by the serial dilution methodology.

1. Place 50 µL diluted saline Buffer onto each of five circles of the slide.2. Using a 50 µL micro pipette add 50 µL serum sample to the drop of saline buffer

in 1st circle.3. Using the same micro pipette, mix the sample with saline by aspirating back &

forth several times. Aspirate 50 µL from 1st circle and transfer to 2nd circle.Repeat the same operation up to 5th circle. Aspirate 50 µL from 5th circle anddiscard. Dilutions obtained as 1/2,1/4, 1/8, 1/16, 1/32

4. Then add 1 drop of RPR antigen suspension (18 µL) to the above circles.Mix androck the slide gently to and fro for 8 minutes; observe the agglutination undergood light source for appearance of carbon particle clumping.

INTERPRETATION

The titre of reagin antibodies is the highest dilution of the test sample which isreactive.

NOTE

1. As with tests based on reagin antibody detection, the RPR syphilis test mayproduce false positive results. These can be linked to diseases such as leprosy,systemic lupus erythoematosus (SLE), infectious mononucleosis, malaria, viralpneumonia. Any positive result must be confirmed by another serological assay(eg.TPHA) Final diagnosis will only be reached after correlation of the resultswith clinical signs.

2. False negative results may be seen in early and latent stages of the disease.3. Qunatitative procedure must be performed to determine the response to

treatment and detect re-infection.4. While dispensing reagents/specimens hold pipette/dropper vertically straight.5. The needle assembly should be thoroughly rinsed with distilled water and air

dried before further use.6. The test cards must not be reused.BIBLIOGRAPHY

1. Caumes E., Janier M. Syphilis, editions techniques. Encyclo. Med Chir (ParisFrance) Maladies Infectiouses. 8-039 -A-10(1994)

2. Drustin L.M. syphilis. Curr OPN. Infect Dis2 : 11-15 (1989)

RPR 50 T, 125 T, 500 T

12204001, 12204002, 12204003


79

Blood GROUPING

INTENDED USE

This reagent is a monoclonal antibody solution intended for determination of red cells

having A antigen on the surface, who are categorized as A group individuals.

- Monoclonal antibody technology

- High avidity and high titre

- Convenient pack sizes

PRINCIPLE

This technique employs the principle of hemagglutination. When the red blood cells

bearing the A antigens are mixed with the Anti-A solution, if agglutination occurs

indicates the presence of A antigen, and the individual belongs to A group and if no

agglutination occurs indicated absence of A antigen.

REAGENT COMPOSITION

Anti A is ready to use reagent prepared from supernatents of mouse hybridoma cell

culters. These reagents contain sodium azide (<0.1%), sodium arsenite (0.02%) and

bovine albumin.


The sealed reagent is stable up to expiry date stated on the vial label, when stored at

2-80C. It is advisable to minimize its time outside the refrigerator between two uses.

SAMPLE

Whole blood sample must be examined with in 48 hrs. Samples should be stored at

2-80C, if not tested immediately. Do not use haemolysed samples.


PLATE TECHNIQUE AT ROOM TEMPERATURE

Bring the reagents to room temperature. Place one drop of Anti-A reagent on a clean

slide, and then add one small drop of whole blood. Mix well with a mixing stick

uniformly. Rock the slide gently back and forth for about 3 minutes while

macroscopically observing the possible appearance of agglutination.

DIRECT METHOD IN A TUBE AT ROOM TEMPERATURE

Prepare a 5% suspension of red blood cells in isotonic saline solution. Using the vial

dropper, place one drop of Anti-A reagent in to labeled test tube. Add a drop of 5 %

RBC suspension. Shake to homogenize the mixture, then centrifuge at 1000 RPM for

60 seconds. Read macroscopically for appearance of any agglutination.

INTERPRETATION

If there is agglutination (ie clumping of red blood cells) the test result indicates the

presence of A antigen.

If there is no visible agglutination, (ie clumping is absent), the test result indicates the

absence of A antigen.

NOTE:

For getting high titre use ABD dilution buffer (code.13601330). The ABD dilution

buffer will be provided on request.

BIBLIOGRAPHY

1. Mannessier, L.; Blood transfusion center Lille France. The use of monoclonal

antibodies as blood grouping reagents: applications, advantages and problems.

Congress of the Italian society for blood transfusion –Rome-June 1992.

2. Blood group serology, Boorman, Dodd and Lincol Churchil Livingstone 6th edition.

3. Human blood groups, Geoff Daniela, 1st edition Blackwell Science, Oxford 1995.

ANTI - A1 x 10 mL

13601010

80

Blood GROUPING

INTENDED USE

This reagent is intended for in vitro determination of ABO group and Rh typing ofhuman red blood cell antigens.


- Highly specific

- Convenient slide and tube formats.


The ABO grouping is defined by both, the presence of ‘A’ and/or ‘B’ antigens on thesurface of the red blood cells, and by simultaneous presence of Anti-A and or Anti-Bantibodies in the serum. An individual has in his serum the antibodies correspondingto the antigens which are not present on his red blood cells.

Based on the principle of hemagglutination, human red blood cells possessing ‘A’ and/or ‘B’ antigen will agglutinate in the presence of corresponding antibody directedtowards the antigen.

Human red blood cells are classified as Rh positive (Rh+) or Rh negative (Rh-) dependingupon the presence or absence of ‘D’ antigen on them. Red blood cells possessing Dantigen will agglutinate in the presence of reagent containing corresponding antibody.

REAGENT COMPOSITION

Anti-A, Anti-B and Anti-D are ready to use reagents prepared from supernatents ofmouse hybridoma cell cultures. These antibodies of immunoglobin class IgM (Anti-D IgM+IgG) are a mixture of several monoclonal antibodies of the same specificitybut having capacity of recognising different epitopes of human red blood cellantigens ‘A’, ‘B’ and ‘D’ (Rh)

Anti-A 1 x 10 mL

Anti-B 1 x 10 mL

Anti-D(IgG+IgM) 1 x 10 mL

These reagents contain sodium azide (<0.1%), sodium arsenite (0.02%) and bovinealbumin.


The sealed reagents are stable up to expiry date stated on the label when storedat 2-80C. It is advisable to minimize its time outside the refrigerator between two uses.

SAMPLE

Blood collected with or without anticoagulant : EDTA, heparin or citrate,

stored beween 20C to 80C upto 48 hours. Haemolysed blood should not be used.



Bring the reagents to room temperature. Place one drop of Anti-A,Anti -B & Anti Dreagent on a clean slide, and then add one small drop of whole blood. Mix well with amixing stick uniformly. Rock the slide gently back and forth for about 3 minutes whilemacroscopically observing the possible appearance of agglutination.


Prepare a 5% suspension of red blood cells in isotonic saline solution. Using the vialdropper, place one drop of Anti-A,Anti -B & Anti D reagent in to labeled test tube.Add a drop of 5 % RBC suspension. Shake to homogenize the mixture, then centrifugeat 1000 RPM for 60 seconds. Read macroscopically for appearance of anyagglutination.

INTERPRETATION

With both the above methods, if there is agglutination (the red blood cells from oneor several clumps) the reaction is positive and the antigen or at least one of theantigens corresponding to the reagent used is present on the red blood cells tested.

ANTI - A, B & D(IgG+IgM)

3 x 10 mL

13601001

If there is no agglutination (red blood cells form a homogenous suspension), thereaction is negative and the antigen is not present on the red blood cells.

In case of anti D agglutination a positive test indicates presence of Rh antigen i.e. Rhpositive. No agglutination or a negative test indicated either presence weaker variantsof Rh antigens like Du or absence of Rh antigen. This needs confirmation by carryingout indirect coomb’s test.

Drying at the periphery or fibrin strands should not be misinterpreted as agglutination.

NOTE:

For getting high titre use ABD dilution buffer.(code.13601330). The ABD dilutionbuffer will be provided on request.

BIBLIOGRAPHY

1. Common Technical Specifications (CTs) for the in vitro diagnosis medical devicesof annex II, list A, of directive 98/79/EC are notified in the official journal of theEuropean communities under document No.C (2202) 1344, with EEA relevance(2202/364/CE)

2. Mannessier .L Blood transfusion center Lille France. The use of monoclonalantibodies as blood grouping reagents: applications, advantages and problems.Congress of the Italian society for blood transfusion –Rome-June 1992.

3. Blood group serology, Boorman, Dodd and Lincoln, Churchill Livingstone, 6th

edition.4. Human blood groups, Geoff Daniels, 1st edition, Blackwell Science, Oxford 1995.


81

Blood GROUPING

INTENDED USE

This reagent is intended for in vitro determination of ABO group and Rh typing ofhuman red blood cell antigens.

Monoclonal antibody technology

Highly specific

Convenient slide and tube formats


The ABO grouping is defined by both, the presence of ‘A’ and/or ‘B’ antigens on thesurface of the red blood cells, and by simultaneous presence of Anti-A and or Anti-Bantibodies in the serum. An individual has in his serum the antibodies correspondingto the antigens which are not present on his red blood cells.

Based on the principle of hemagglutination, human red blood cells possessing ‘A’ and/or ‘B’ antigen will agglutinate in the presence of corresponding antibody directedtowards the antigen.

Human red blood cells are classified as Rh positive (Rh+) or Rh negative (Rh-) dependingupon the presence or absence of ‘D’ antigen on them. Red blood cells possessing Dantigen will agglutinate in the presence of reagent containing corresponding antibody.

REAGENT COMPOSITION

Anti-A, Anti-B and Anti-D are ready to use reagents prepared from supernatents ofmouse hybridoma cell cultures. These antibodies of immunoglobin class IgM (Anti-DIgM) are a mixture of several monoclonal antibodies of the same specificity but havingcapacity of recognising different epitopes of human red blood cell antigens ‘A’, ‘B’ and‘D’ (Rh)

Anti – A 1 x 10 mL

Anti – B 1 x 10 mL

Anti – D (IgM) 1 x 10 mL

These reagents contain sodium azide (<0.1%), sodium arsenite (0.02%) and bovinealbumin.


The sealed reagents are stable up to expiry date stated on the label when stored at 2-80C. It is advisable to minimize its time outside the refrigerator and to avoid leaving itat room temperature between two uses.

SAMPLE

Blood collected with or without anticoagulant : EDTA, heparin or citrate, stored beween20C to 80C upto 48 hours. Haemolysed blood should not be used.


Plate technique at room temperature

On a rigorously clean plate, using the vial dropper, apply one drop of each reagent.Take one drop of blood and apply next to each drop of reagent, taking care not tocreate contact between the drops. Mix the blood and reagent using a spiral movementwith the end of the stirrer so as to create a regular lozenge of diameter 2 to 3 cm.Incubate the plate at room temperature and with out stirring for 30 seconds. Holdthe plate and give it a rolling movement for 3 minutes while macroscopically observingthe possible appearance of agglutinates. Read reaction immediately.

Direct method in a tube at room temperature

Prepare a 5% suspension of red blood cells in isotonic saline solution. Using the vialdropper, transfer a drop reagent Anti-A, Anti-B and Anti-D into correspondingly labeledtubes. Add a drop of red blood cell suspension. Shake to homogenize the mixture,then centrifuge at 1000 rpm for 1 minutes. Read macroscopically while gently shakingthe tubes. Note the appearance of any agglutinates.

INTERPRETATION

With both the above methods, if there is agglutination (the red blood cells from oneor several clumps) the reaction is positive and the antigen or at least one of theantigens corresponding to the reagent used is present on the red blood cells tested.

If there is no agglutination (red blood cells reform a homogenous suspension), thereaction is negative and the antigen is not present on the red blood cells.

In case of anti D agglutination is a positive test indicates presence of Rh antigen i.e.Rh positive. If there is no agglutination (i.e. clumbing is absent), it indicates eitherpresence of weaker varients of Rh antigens like Du or absence of Rh antigens. It ispossible to use Anti TOTEM in an andirect coomb’s test if weak / or partial antigens Dare to be detected.

Drying at the periphery or fibrin strands should not be misinterpreted as agglutination.

BIBLIOGRAPHY

1. Common Techanical Specifications (CTs) for the in vitro diagnosis medical devicesof annex II, list A, of directive 98/79/EC are notified in the official journal of theEuropean communities under document No.C (2202) 1344, with EEA relevance(2202/364/CE)

2. Mannessier, .L ; Blood transfusion center Lille France. The use of monoclonalantibodies as blood grouping reagents: applications, advantages and problems.Congress of the Italian society for blood transfusion –Rome-June 1992.



ANTI - A, B & D(IgM)

3 x 10 mL

13601013

82

Blood GROUPING

INTENDED USE

This reagent is a monoclonal antibody solution intended for determination of red cells

having B antigen on the surface, who are categorized as B group individuals.




PRINCIPLE

This technique employs the principle of hemagglutination. When the red blood cells

bearing the B antigens are mixed with the Anti-B solution, if agglutination occurs

indicates the presence of B antigen, and the individual belongs to B group and if no

agglutination occurs indicated absence of B antigen.

REAGENT COMPOSITION

Anti B is ready to use reagent prepared from supernatents of mouse hybridoma cell

culters. These reagents contain sodium azide (<0.1%), sodium arsenite (0.02%) and

bovine albumin.


The sealed reagent is stable up to expiry date stated on the vial label, when stored at

2-80C. It is advisable to minimize its time outside the refrigerator between two uses.

SAMPLE

Whole blood sample must be examined with in 48 hrs. Samples should be stored at

2-80C, if not tested immediately. Do not use haemolysed samples.



Bring the reagents to room temperature. Place one drop of Anti-B reagent on a clean

slide, and then add one small drop of whole blood. Mix well with a mixing stick

uniformly. Rock the slide gently back and forth for about 3 minutes while

macroscopically observing the possible appearance of agglutination.


Prepare a 5% suspension of red blood cells in isotonic saline solution. Using the vial

dropper, place one drop of Anti-B reagent in to labeled test tube. Add a drop of 5%

RBC suspension. Shake to homogenize the mixture, then centrifuge at 1000 RPM for

60 seconds. Read macroscopically for appearance of any agglutination.

INTERPRETATION

If there is agglutination (ie clumping of red blood cells) the test result indicates the

presence of B antigen.

If there is no visible agglutination, (ie clumping is absent), the test result indicates the

absence of B antigen.

NOTE:

For getting high titre use ABD dilution buffer (code.13601330). The ABD dilution

buffer will be provided on request.

BIBLIOGRAPHY

1. Mannessier, L.; Blood transfusion center Lille France. The use of monoclonal

antibodies as blood grouping reagents: applications, advantages and problems.

Congress of the Italian society for blood transfusion –Rome-June 1992.

2. Blood group serology, Boorman, Dodd and Lincol Churchil Livingstone 6th edition.

3. Human blood groups, Geoff Daniela, 1st edition Blackwell Science, Oxford 1995.

ANTI - B 1 x 10 mL

13601009


83

Blood GROUPING

INTENDED USE

This reagent is intended for in vitro determination of presence or absence of Rho(D) antigen on human red blood cells.




- Detects most variants of D antigen


Human erythrocytes having Rho Antigen (D-Ag) will cause agglutination is thepresence of Anti-D. Antibodies present in the reagent Human red blood cells areclassified as Rho(D) positive or Rho (D) negative depending upon the presence orabsence of D(Rho) antigen on them. All negative test results should be furtherconfirmed by coomb’s test.

REAGENT COMPOSITION

The Anti –D is a monoclonal blend (IgG+IgM) antibodies derived from thesupernatant of Hybridomas of murine (or) human origin. These reagents containsodium azide (<0.1%) sodium arsenite (0.02%) and bovine albumin.


The sealed reagents are stable up to expiry date stated on the label when storedat 2-80C. It is advisable to minimize its time outside the refrigerator and to avoidleaving it at room temperature between two uses.

SAMPLE

Whole blood samples must be examined within 48 hrs. Samples should be storedat 2-80C, if not tested immediately. Do not use haemolysed samples.


A) PLATE TECHNIQUE AT ROOM TEMPERATURE

Bring the reagents to room temperature. Place one drop of Anti-D (IgG+IgM) reagenton a clean slide, then add one small drop of whole blood. Mix well with a mixingstick uniformly. Rock the slide gently back and forth for about 3 minutes whilemacroscopically observing the possible appearance of agglutinates.

B) DIRECT METHOD IN A TUBE AT ROOM TEMPERATURE

Prepare a 5% suspension of red blood cells in isotonic saline solution. Using thevial dropper, transfer a drop reagent Anti- D(IgG+IgM)reagent in to a labelled testtube. Add a drop of red blood cell suspension. Shake to homogenize the mixture,then centrifuge at 1000 rpm for one minute. Read macroscopically while gentlyshaking the tubes so as to detach the RBC pellets. Note the appearance of anyagglutinates.

INTERPRETATION

If there is agglutination (i.e. clumping red blood cells) while using directhemagglutination method on a plate or in a tube, the test result indicates thepresence of D antigen (Rh positive)

If there is no agglutination, while using direct hemagglutination method on a plateor in a tube, the test result indicates the absence of D antigen (Rh negative)

The presence or absence of weak and / or partial D antigen, should be confirmedby carrying out Indirect Antiglobulin Test (IAT)

Drying at the periphery or fibrin strands should not be misinterpreted asagglutination.

NOTES

1. The reactions must be read immediately after centrifuging and re-suspending.2. Anti D (IgG+IgM) have the special feature of recognizing certain rare antigen

motives of type RH33 (RoHar) and may thus yield discordant reactions withpolyclonal reagents that recognize them little or not at all.

3. For specific identification of partial D antigen, the use of any 3rd party kit forthis purpose is recommended.

4. A false positive reaction may occur if the subject tested has cold agglutinatinins5. Anti D (IgG+IgM) cannot ensure the recognition of all weak or variant subjects,

due to the variability of antigen motifs.6. Certain discordances (negative reaction for the direct hemagglutination method

and positive reaction for the indirect antiglobulin method) may occur with AntiD (IgG+IgM). A weak and/or partial D antigen may be suspected.

7. A false –positive reaction may occur, when Anti D (IgG+IgM) is used in theindirect antiglobulin method (IAT), if the RBC from the test subject shows apositive reaction in the direct antiglobulin test (DAT)

8. A negative reaction obtained in an indirect antiglobulin test can be validatedwith IgG-sensitized red blood cells (not provided with this kit)

CLONAL SPECIFICITY OF ANTI D (IgG+IgM)

D

IIIa D D D D D D D HMi

D IIIb IVa IVb Va VI VII FR BT RoHor

Clones Type II IIIc (RH33)

P3X61 IgM + + + + + - + +/- + + +

P3X212 IgM - + - - + + + + - - +

23B10

P3X290 IgG + + +/- - + +/- + + - - +

P3X35 IgG + + + + - - + - - - +

The table gives the specificity of clones used in Anti D (IgG+IgM) reagent againstvariants D Antigen. The intensity of reactions obtained with Anti D (IgG+IgM) mayvary as a function of the number of antigen sites present on the test red blood cells.

BIBLIOGRAPHY

1. Mannessier, L.; Blood transfusion center Lille France. The use of monoclonalantibodies as blood grouping reagents: applications, advantages and problems.Congress of the Italian society for blood transfusion –Rome-June 1992.



ANTI - D(IgG+IgM)

1 x 10 mL

13601003


9. For getting high titre use ABD dilution buffer (code.13601330) . The ABD dilutionbuffer will be provided on request.

84

Blood GROUPING

INTENDED USE

This reagent is intended for in vitro determination of presence or absence of Rho (D)antigen on human red blood cells.

- Monoclonal IgM Antibodies



PRINCIPLE

The test procedure recommended for the use of this antisera are based upon theagglutination (clumping) of red blood cells carrying the D antigen in the presence ofan IgM Anti-D antibody. All negative test results should be further confirmed by Dutest.

REAGENT COMPOSITION

The reagents contain monoclonal antibodies. The monoclonal antibodies are derivedfrom the supernantants of in vitro cultures of hybridomas of murine origin. Thesereagents contain sodium azide (<0.1%) sodium arsenite (0.02%) and bovine albumin.


The reagents must be stored between +2oC and +80C. It is advisable to minimize itstime outside the refrigerator and to avoid leaving it at room temperature between twouses.

SAMPLE

Blood collected in anticoagulant : EDTA, heparin or citrated in a stoppered sterile tube,stored between 2-80C. At the time of the test, centrifuge the blood sample at 1200 gfor 3 minutes. Do not use hemolysed samples


A) PLATE TECHNIQUE AT ROOM TEMPERATURE

Bring the reagents to room temperature. Place one drop of Anti-D (IgM) reagent on aclean slide, then add one small drop of whole blood. Mix well with a mixing stickuniformly. Rock the slide gently back and forth for about 3 minutes whilemacroscopically observing the possible appearance of agglutination.

B) DIRECT METHOD IN A TUBE AT ROOM TEMPERATURE

Prepare a 5% suspension of red blood cells in isotonic saline solution. Using the vialdropper, transfer a drop reagent Anti- D(IgM)reagent in to a labelled test tube. Add adrop of red blood cell suspension. Shake to homogenize the mixture, then centrifugeat 1000 rpm for one minute. Read macroscopically while gently shaking the tubes soas to detach the RBC pellets. Note the appearance of any agglutination.

INTERPRETATION

If there is agglutination (ie, clumping red blood cells) with Anti-D(IgM) antigen D ispresent.

If there is no agglutination (ie, clumping is absent) it is possible to use Anti D (IgG)in a coomb’s test to rule out the presence of weak and partial D antigens.

NOTES

1. The reactions must be read immediately after centrifuging and resuspending.2. A false positive reaction may occur if the subject tested has cold agglutinins.3. Anti D (IgM) cannot ensure the recognition of weak or variant subjects, due to

the variability of antigen motifs.4. For getting high titre use ABD dilution buffer (code.13601330) . The ABD dilutionbuffer will be provided on request.

BIBLIOGRAPHY

1. Betermieux, C., Beolet, M. , Keyser, L.; A new strategy for D phenotyping withTOTEM multimonoclonal ANTI –D reagent, XX111rd I.S.B.T. Congress, July 1994.

2. Aramburu, E., Rabasa, P. , Esquiroz, R., Galarretat, B. Valoracion de un antisureoanti-D IgM-IgG monoclonal (DIAGAST) en donantes de sangre concentrationexpresividaddedli del anti geno. Hematology congress, Madrid, October 1990.

3. Mannessier, L. Blood transfusion center, Lille, France. The use of monoclonalantibodies as blood grouping reagents: applications, advantages and problems.Congress of the Italian society for blood transfusion ,Rome,June 199

ANTI - D(IgM)

1 x 10 mL

13601015

85

INTENDED USE

This reagent is intended for the in vitro determination of Prothrombin Time in citratedplasma.


The Prothrombin Time is an indicator of the extrinsic blood coagulation mechanism.Deficiencies of Prothrombin and co factors V, VII and X give rise to a prolongedclotting time.

Presence of high levels of Heparin in the blood sample hypofibrinogenemia alsoattribute for prolonged time.

Common causes prolonged one stage prothrombin time are:

1. Therapy with coumarin or indanedione drugs.2. Obstructive jaundice3. Hemorrhagic disease of the new born4. Liver disease.Less common causes:1. Heparin therapy2. Loss of clotting proteins from blood via kidneys in renal disease.

Eg. Nephrotic syndrome3. Congenital deficiency of clotting factor(s)4. Fibrinogen deficiency5. Vitamin K deficiency

PRINCIPLE

Tissue thromboplastin in the presence of Ca++ activates extrinsic pathway of humanblood coagulation cascade. Activation time is proportional to the concentration ofindividual clotting factors taking part in the coagulation cascade. This assists inestimating cause and extent of haemorrhagic disorder.

When thromboplastin reagent is added to citrated plasma, clotting cascade isinitiated forming gel clot. The time required for clot formation would be prolongedif there is deficiency of factor (s) activity in the extrinsic pathway of the coagulationcycle.

REAGENT COMPOSITION

PROTHROMBIN TIME (PT)

TF (Rabbit origin)

Ca++

Antimicrobials

Stabilizers

PACK CONTENTS

PT Reagent 2 x 4 mL

Buffered tri sodium citrate (0.109M) 3.2% 1 x 10 mL



Do not freeze.

NORMAL RANGE

Clotting time : 10 - 15 sec

Prothrombin ratio : 1 ± 0.15 (depends on methodology)

INR value : 0.8 -1.5

The clotting time of abnormal plasma will depend on the International SensitivityIndex (ISI) of the reagent lot. Normal range should be established in the user’slaboratory.

SAMPLE COLLECTION PREPARATION

Citrated plasma.

PT (S.L) 2 x 4 mL

12601003

Mix gently, 9 parts of blood in a plastic tube or siliconized glass tube containing 1part of 3.2% tri sodium citrate solution (0.109 M). Centrifuge immediately for 15min at 3000 rpm to obtain platelet poor plasma. Transfer supernatant plasma in asiliconized glass tube or plastic tube immediately, do not disturb buffy coat whilecollecting supernatant plasma. For best results the test should be done within 2hours of blood collection.

TEST PROCEDURE

The PT for each sample should be determined at least twice. This procedure pertainsto manual or semi-automated coagulation systems. Refer to your instrumentmanual for more detailed instrument specific instruction.

Manual Method

1. Gently swirl the reagent vials before use. Do not shake.2. Dispense from the vial enough PT Reagent for immediate use, into a thoroughly

clean dry test tube.3. Pre-warm the dispensed PT Reagent to 370C for 10 minutes.4. Pipette 100 µL of plasma into test cuvette at 370C, and incubate for

3 minutes.5. Add forcibly 200 µL pre-warmed PT reagent into the test cuvette.6. Start timer simultaneously and record the clotting time in seconds.

CALCULATION

The ISI as well as nature of preparation varies worldwide, it is recommended toexpress results of the coagulation assays in terms of INR and this can be derivedfrom the accompanying INR conversion table or calculated from the belowmentioned relationship

(International Normalized Ratio) INR = RISI where,

Mean PT of the patients plasma in sec

R(prothrombin ratio) = -----------------------------------------------------

MNPT (Mean Normal PT) for the reagent

MNPT: Each laboratory must establish its own MNPT for each lot of reagent andinstrument used. Usually plasma from at least 20 normal healthy individual shouldbe used to establish the MNPT. ISI value of the reagent is mentioned on the viallabel.

PRECAUTION

Venous blood should be directly drawn into the tube containing anticoagulant.

Ensure that the sample is free of microclots.

Separate plasma immediately by centrifugation after collection of blood.The test should be done within two hours of blood collection.

Plasma must be stored in siliconized glass tubes or plastic containers.

Avoid turbid, lipemic or hemolyzed samples.

Use clean dry micropipette tips and plasticware to dispense the reagent.

Close reagent vial and replace immediately to 2-80C after dispensing.

GUARANTEE

This product is guaranteed to perform as described on label and package insert.The manufacturer disclaims any warranty of use and sale for any other purpose.

BIBLIOGRAPHY

1. Dacie, J.V., Lewis, S.M.; Practical hematology. 19842. R Biggs, R, Mcfarlane, R. G; Human blood coagulation and its disorders 19633. Burtis, et al. Tietz: Text book of Clinical Chemistry AACC 19994. Roadck, B. F.; Diagnostic Hematology, clinical principles and applications 2nd

edition.5. John Bernad Henry: Clinical Diagnosis and management by laboratory methods

20th edition.6. Data on file of Agappe Diagnostics Ltd. Kerala.


86

INTENDED USE

This reagent is intended for the in vitro determination of Activated PartialThromboplastin Time (APTT) in citrated plasma.


The activated partial thromboplastin time (APTT) is used as a general screening testfor the detection of coagulation abnormalities in the intrinsic pathway. APTT is sensitiveto deficiencies or abnormalities of factors VIII, IX, XI, XII, X, V, II and I. APTT is alsosensitive to inhibitors of blood coagulation such as lupus inhibitor and fibrin/fibrinogen degradation products. APTT is the most widely used method for monitoringintravenous heparin anticoagulation therapy.

PRINCIPLE

In presence of Calcium ions cephaloplastin activates coagulation factors of intrinsicpathway in plasma leading to clot formation. Clotting time is proportional to theconcentration of factors VIII, IX, XI and XII as well as common pathway factors II, V,and X. As the reagent is prepared using one single species rabbit brain, it has therequired sensitivity to be used in heparin assays, also has better sensitivity for factorsVIII & LA.

PACK CONTENTS

APTT Reagent 1 2 x 4 mL

Calcium chloride solution 0.020 M/L

APTT Reagent 2 2 x 4 mL

Rabbit brain cephalin

Ellagic acid activator

Buffer

Stabilizers and preservatives

Tri sodium citrate (0.109 M/L) 3.2% 1 x 10 mL



Do not freeze.

NORMAL RANGE

The normal values are between 21-38 seconds (at 3 minutes activation). The normaltime depends on the method, activation time, instrument etc. and must be determinedin each laboratory.

PRECAUTION

Venous blood should be directly drawn into the tube containing anticoagulant.

Ensure that the sample is free of microclots.

Separate plasma immediately by centrifugation after collection of blood.

The test should be done preferably within two hours of blood collection.

Plasma must be stored in siliconized glass tubes or plastic containers.

Avoid turbid, lipemic or hemolyzed samples.

Use clean dry micropipette tips and plasticware to dispense the reagent.

Mix the reagent (by gentle swirling) before use.

Close reagent vial and replace immediately to 2-80C after dispensing.

SAMPLE COLLECTION & PREPARATION

Citrated plasma.

Mix gently, 9 parts of blood in a plastic tube or siliconzied glass tube containing 1part of 3.2% tri-sodium citrate solution (0.109 M/L). Centrifuge immediately for 15minutes at 3000 rpm to obtain platelet poor plasma. Transfer supernatant plasma ina siliconized glass tube or plastic tube immediately; do not disturb the buffy coat whilecollecting supernatant plasma. The test should be done preferably within 2 hours ofblood collection.

TEST PROCEDURE

APTT of each sample should be determined at each level twice. This procedure pertainsto manual or semi-automated coagulation systems. Refer to your instrument manualfor more detailed instrument specific instructions

APTT 2 x 4 mL

12602001

Bring contents of the vial to room temperature and then swirl gently to mix to ahomogenized suspension. Keep the reagents at 370C for 10 minutes prior to use.

1. Gently swirl the reagent vials before use. Do not shake.2. Pre-warm enough volume of Reagent1 (CaCl

2) for immediate use, in a clean & dry

plastic test tube at 370C.3. Pipette 100 µL of test plasma or control in to a test cuvette at 370C.4. Pipette 100 µL of the pre-warmed reagent 2 (APTT Reagent) in to the test cuvette.5. Mix well and incubate at 370C for 3 minutes.6. Forcibly pipette 100 µL of pre warmed Reagent1 (CaCl

2) into the test cuvette.

7. Start a timer simultaneously and record the clotting time in seconds.Calibration curve for the determination of Heparin concentration:

Dilute Heparin (as used for treatment) with physiological saline to concentration of10 IU/mL.

Mix 0.2 mL of 10 IU/mL diluted heparin with 1.8 mL of FNP (Fresh Normal Plasma) toyield heparin standard of 1 IU/mL concentration.

Dilute the Heparin standard as prepared above (1 IU/ml) with FNP as follows.

Test Tube 1 2 3 4 5 6 7

Heparin std.(1IU/mL) 0.5 0.4 0.3 0.2 0.1 0.1 -

FNP in mL - 0.1 0.2 0.3 0.4 0.9 0.5

Heparin con.(IU/mL) 1 0.8 0.6 0.4 0.2 0.1 0

Procedure: Manual method to estimate Heparin concentration in the plasma/sample.

. Pipette 0.1mL of each Heparin dilution into clean test tubes.

. Add 0.1 mL APTT Reagent (pre-warmed at 370C). Mix and incubate for exactly3 minutes at 370C.

. Add. 0.1 mL Pre-warmed Calcium chloride (0.0290 M/L) and simultaneously startstopwatch.

. Observe clot formation carefully and note the time at the appearance of firstfibrin web.

. Plot mean of double determination in seconds against each heparinconcentration on heparin calibration graph paper provided.

. Connect points in a straight line.*Plot clotting time of sample on the calibration curve and read heparinconcentration in IU/mL

Note: Laboratories using coagulometers should follow instructions (sequence) ofthe coagulometer manufacturer.

Warranty: The product is designed to perform as described in the pack insert. Themanufacturer disclaims any implied warranty of use and sale for any other purpose.

CALCULATION

The result of APTT test can be reported directly in seconds.

BIBLIOGRAPHY

1. Dacie, J.V., Lewis, S.M.; Practical hematology. 19842. R Biggs, R, Mcfarlane, R. G; Human blood coagulation and its disorders 19633. Burtis, et al. Tietz: Text book of Clinical Chemistry AACC 19994. Roadck, B. F; Diagnostic Hematology, clinical principles and applications 2nd

edition.5. John Bernad Henry: Clinical Diagnosis and management by laboratory methods

20th edition.6. Data on file of Agappe Diagnostics Ltd. Kerala.


87

Haemo CHEK

INTENDED USE

Diluent is intended for counting and sizing of blood cells in MINDRAY- 3 partDifferential Hematology Analyzers of BC Series. It is an azide free filtered isotonicsolution.

REAGENT COMPOSITION

Sodium chloride 3 - 5.5 g/L

Sodium sulphate anhydrous 7.5 - 11.5g/L

Buffering agents 1- 3 g/L

Anti fungal & anti bacterial Agent 0.8 - 2.5 g/L


The sealed reagents are stable up to the expiry date stated on the label, when storedat 150C to 300C.

ONCE OPENED STABLE FOR 60 DAYS.

DIRECTIONS FOR USE

Follow the procedures given in the operation manual of Mindray 3 part differentialHematology Analyzer.

PRECAUTIONS AND WARNING

1. For in vitro diagnostic use only.2. Do not pipette by mouth.3. Do not inhale the reagent.4. If the reagent comes in contact with, eye or skin wash off immediately with large

quantity of water and seek medical attention immediately5. Do not use the reagent in any procedures other than those described herein6. Do not use containers and other materials in the package for any purpose other

than those described herein7. When discarding the reagents dispose of them according to local regulations.8. Take normal precautions required for handling laboratory reagents.

DILUENT1 x 10 Litre, 1 x 20 Litre

14001001, 14001002


88

Haemo CHEK

INTENDED USE

EZ cleaner is intended for daily cleaning of MINDRAY- 3 part Differential HematologyAnalyzers of BC Series.

REAGENT COMPOSITION

Proteolytic enzyme 3.0 - 10 g/L

Substrate 0.3 - 1.5 g/L

Sodium chloride 3.0 - 5.0 g/L

Buffering agents 1.0 - 4.0 g/L

Anti fungal & anti bacterial agent 0.5 -2.5 g/L



Discard the reagent if kept opened for more than 30 days.

DIRECTIONS FOR USE

Follow the procedures given in the operation manual of Mindray 3 part differentialhematology analyzer.


1. For in vitro diagnostic use only.2. Do not pipette by mouth.3. Do not inhale the reagent.4. If the reagent comes in contact with eye or skin, wash off immediately with large

amount of water and seek medical attention immediately5. Do not use the reagent described above in any procedures other than those

described herein.6. Do not use containers and other materials in the package for any purpose other

than those described herein.7. When discarding the reagents dispose of them according to local regulations.8. Attend to the normal precautions required for handling all laboratory reagents.

E-Z CLEANER4 x 50 mL

14002001


89

Haemo CHEK

INTENDED USE

Lyse is intended for lysing of cells for WBC Count and Hemoglobin measurementin MINDRAY-3 part Differential Hematology Analyzers of BC Series.

REAGENT COMPOSITION

Quaternary ammonium salts - < 50 g/L

Non-ionic surfactant - < 15 g/L

2-Propanol - 0.1 - 1.5mL/L



ONCE OPENED STABLE FOR 60 DAYS

DIRECTIONS FOR USE

Follow the procedures given in the operation manual of Mindray 3 part differentialhematology analyzer.


1. For in vitro diagnostic use only.2. Do not pipette by mouth.3. Do not inhale the reagent4. If the reagent comes in contact with eye or skin, wash off immediately with large

quantity of water and seek medical attention immediately5. Do not use the reagent described above in any procedures other than those

described herein.6. Do not use containers and other materials in the package for any purpose other

than those described herein.7. When discarding the reagents dispose of them according to local regulations.8. Attend to the normal precautions required for handling all laboratory reagents.

LYSE1x500 mL/2x500 mL

14003001/14003002


90

Haemo CHEK

INTENDED USE

Probe cleaner is intended for periodical cleaning of sample probe inMINDRAY-3 partDifferential Hematology Analyzers of BC Series.

REAGENT COMPOSITION

Surfactant > 2.0 g/L

Sodium hypochlorite > 100 g/L

Sodium chloride > 100 g/L



DIRECTIONS FOR USE




amount of water and seek medical attention immediately5. Do not use the reagent described above in any procedures other than those

described here in.6. Do not use containers and other materials in the package for any purpose other

than those described here in.7. When discarding the reagents dispose of them according to local regulations8. Attend to the normal precautions required for handling all laboratory reagents.

PROBE CLEANER4 x 50 mL

14004001


91

Haemo CHEK

INTENDED USE

Rinse is intended for cleaning the volumetric tubes of MINDRAY-3 part DifferentialHematology Analyzers of BC Series.

It is an azide free filtered solution.

REAGENT COMPOSITION

Sodium chloride 3.0 - 5.5 g/L

Sodium sulphate anhydrous 7.5 - 11.5 g/L

Buffering agents 1.0 - 3.0 g/L

Non ionic surfactant 5.0 - 8.0 g/L

Anti fungal & anti bacterial agent 0.8 - 2.5 g/L



ONCE OPENED THE REAGENT IS STABLE FOR 60 DAYS

DIRECTIONS FOR USE




quantity of water and seek medical attention immediately.5. Do not use the reagent described above in any procedures other than those

described here in.6. Do not use containers and other materials in the package for any purpose

other than those described here in.7. When discarding the reagents dispose of them according to local regulations.8. Attend to the normal precautions required for handling all laboratory reagents.

RINSE1 x 10 Litre, 1 x 20 Litre

14005001, 14005002


92

INTENDED USE

This reagent is intended for the preparation of calibration curve for ASO estimation.

REAGENT COMPOSITION

ASO calibrator is prepared by diluting human plasma which contains high levels ofAnti Streptolysin – O (ASO) with physiological saline containing 1%(w/v) bovineserum albumin.


The sealed vials are stable up to expiry date if kept at 2-80C, protected from light.

PREPARATION AND STABILITY OF CALIBRATOR

Calibrator is ready to use and concentration is mentioned on the vial.

PRECAUTION

To avoid contamination, use clean laboratory wares. Close the vials immediatelyafter use. Protect from light and heat. Improper handling or storage of thecalibrators can affect results.

WARNING

Human source material. Each donor unit although tested negative for HbsAg, HCV,HIV(1&2) treat as potentially infectious material

Reagents containing sodium azide must be handled with care.

BIBLIOGRAPHY

1. Galuin, J.Pet al., Particle enhanced photometric immunoaasay system2. Singer, J. M.et al., The latex fixation test Application to the serologic diagnosis

of rheumatoid arthritis, Amer. J. med., 21, 888(1956)

ASO CALIBRATOR1 x 1 mL

11615003

ADL/V.01/APR 2012

93

INTENDED USE

This reagent is intended for the preparation of calibration curve for microalbuminestimation.

REAGENT COMPOSITION

A dilution of defibrinated human albumin solution with phosphate buffered saline,liquid stabilized filtered through 0.2m filter.

Sodium azide – 0.095%


The sealed vials are stable up to expiry date at 2-80C, protected from light.


Calibrator is stable liquid and the concentration will be mentioned on the vial.Generate a calibration curve by diluting the calibrator as follows:


Prepare the following calibrator dilution using NaCI as diluent. Multiply theconcentration of the microalbumin calibrator by the corresponding factors stated inthe table below to obtain the microalbumin concentration of each dilution.


Cali. (µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Opened vials are stable up to 6 weeks when stored at 2-80C.

PRECAUTION


WARNING

Human source material. Each plasma donor unit although tested negative for HbsAg,HCV, HIV(1&2) treat as potentially infectious material.


BIBLIOGRAPHY

Mount, N.J. Clin, Pathology, 22, 12(1986)

MICROALBUMIN CALIBRATOR1 x 1 mL

11618003

ADL/V.01/APR 2012

94

INTENDED USE

This reagent is intended for the preparation of calibration curve for CRP estimation.

REAGENT COMPOSITION

CRP calibrator is prepared by diluting defibrinated human plasma with phosphatebuffered saline.


The sealed vials are stable up to expiry date when kept at 2-80C, protected from light.


Calibrator is stable liquid and concentration will be mentioned on the vial. Generate acalibration curve by diluting the calibrator serially.

Preparation of calibration Curve:

Prepare the following calibrator dilution using NaCI as diluent. Multiply theconcentration of the CRP calibrator by the corresponding factors stated in the tablebelow to obtain the CRP concentration of each dilution.


Calib.(µL) - 10 10 20 50 100

Saline(µL) 100 150 70 60 50 -

Dil. Factor 0 0.0625 0.125 0.25 0.5 1.0

PRECAUTION

To avoid contamination, use clean laboratory wares. Close the vials immediately afteruse. Protect from light and heat. Improper handling or storage of the calibrators canaffect results.

WARNING

Human source material. Each donor unit although tested negative for HbsAg, HCV,HIV(1&2) treat as potentially infectious material.


BIBLIOGRAPHY

1. Tillett. W.et al: Serological reactions in pneumonia with a non-protein somaticfraction of pneumoccous.J. exp. Med.. 52,561(1930)

2. Ziegenhagen G,. Drahovshy D. Klimishe Bedeutung desc-Reakriven protein. MedKlin 1983; 78:45-50

CRP CALIBRATOR1 x 2 mL

11616001

ADL/V.01/APR 2012

95

INTENDED USE

This reagent is intended for the preparation of calibration curve for Lp(a) estimation.

REAGENT COMPOSITION

Lyophilized human serum.


The sealed vials are stable up to the expiry date at 2-80C, protected from light.


Remove the vial cap carefully and add exactly 1.0 mL of deionized water. After leavingit for more than 10 minutes, shake it gently until the content is completely dissolved.The reconstituted calibrator is stable for 7 days at 2-80C when protected from lightand heat.


Prepare the following calibrator dilution using NaCI as diluent. Multiply theconcentration of the Lp(a) calibrator by the corresponding factors stated in the tablebelow to obtain the Lp(a) concentration of each dilution.


Calib.(µL) - 10 10 25 50 100

Saline(µL) 100 150 70 75 50 -

Dil. Factor 0 0.0625 0.125 0.25 0.5 1.0

PRECAUTION


WARNING


BIBLIOGRAPHY

1. Utermann, G.et al. Lp(a) Glycoprotein phenotypes. Inheritance andrelation to Lp(a) Lipoprotein concentration in plasma.

2. J.Clin Invest., 80, 458(1987) MClea, J . W.et al. C DNA sequence ofhuman apo lipoprotein (a) is homologous, nature, 300, 132(1987)

Lp(a) CALIBRATOR1 x 1 mL

11619001

ADL/V.01/APR 2012

96

INTENDED USE

This reagent is intended for the preparation of reference curves for quantitativeimmunochemical determination of protein in human serum.

REAGENT COMPOSITION

Protein multicalibrator is a defibrinated human plasma, liquid stabilized and filteredthrough 0.2 m filter.

Sodium azide – 0.05%


The sealed vials are stable up to expiry date at 2-80C, if protected from light.


This protein multicalibrator should be treated similar to sample/control and the testneeds to be performed as per the procedure of the respective reagent.

Opened vials are stable up to 6 weeks when stored at 2-80C.

PRECAUTION


WARNING



BIBLIOGRAPHY

1. Dati, F. et al. Med. 13,87 (1889)2. Poulik, M.D. and klesis in The plasma proteinso, vol.2 second

edition.F.W. Putman Acedemic press, New York, PP 52-108

PROTEIN MULTICALIBRATOR1 x 1 mL

11614004

ADL/V.01/APR 2012

97

INTENDED USE

Multicalibrator is a calibration serum intended for calibration of clinical chemistryassays suitable for manual procedure and automated analyzers.

DESCRIPTION

Multicalibrator is lypholized calibrator based on human serum. The concentrationand activities of calibrator components have been chosen so as to ensure optimumcalibration of automated analyzers. The chemical and biological componentsincluded in this product were chosen to act in the serum matrix in a similar mannerto the corresponding components found in human serum.

REAGENT COMPOSITION

Multicalibrator 5 x 3mL

Lyophilized human serum with additives

Bacteriostatic agents and stabilizers.

(The concentration/activities of each calibrator components are Lot Specific. Pleaserefer to the attached value sheet).

PREPARATION

1. Carefully open one multicalibrator vial.2. Remove the cap and carefully lift the rubber stopper without removing it

completely, allowing air to enter the vial through the groove of the lower partof the stopper.

3. Remove the rubber stopper, avoiding any loss of the lyophilized material.4. Carefully pipette exactly the same amount of deionized water as mentioned

on the vial label, into the opened vial5. Replace the rubber stopper carefully.6. Dissolve the contents completely by occasional gentle swirling (within 30

minutes) (avoiding froth formation). DO NOT SHAKE7. Let it stand at room temperature and away from light for 10 minutes.8. Lyophilizate should be completely dissolved before use.


Store at 2-8oC, away from light. Unopened vial is stable until expiry date stated onthe vial label.

Store the reconstituted calibrator vial tightly capped and protected from light whennot in use.

The components of the reconstituted calibrator is stable for,

At least 8 hours at 15-250C

At least 2 days at 2-80C

At least 30 days at -20oC.

Aliquot and freeze once only.

Bilirubin in the reconstituted calibrator (when stored in dark) is stable for;

At least 3 hrs at 15-250C

At least 8 hours at 2-80C

At least 14 days at -20oC.

Aliquot and freeze once only.

Acid phosphate in the reconstituted calibrator (when stored in dark) is stable for;

At least 4 hours 15-250C

At least 1 day at 2-80C

DIRECTIONS FOR USE

Multicalibrator is to be used in accordance with the directions accompanying thereagents or kits referring to the same method, and following the technical datasheet of the reagent in use. While following these directions, multicalibrator hasto be handled as patient serum.

ASSIGNED VALUES AND TRACEABILITY

The assigned values were determined using the methods mentioned in the valuesheet. Determinations were performed under standardized conditions onautomated analyzers using Agappe reagents and Master Calibrator (SRM/NIST whereever applicable).

The calibrator value specified is the median of all values obtained from separateparticipating laboratories in several independent series. Multicalibrator values aretraceable to reference materials (SRM/NIST) where applicable.


1. For in vitro diagnostic use only2. Do not pipette by mouth3. If the multicalibrator comes in contact with, eye or skin, immediately wash off

with large quantity of water, and seek medical attention immediately.4. Do not use containers and other materials in the package for any purpose other

than those described herein.5. When discarding the reagents dispose of them according to local regulations6. Attend to the normal precautions required for handling laboratory reagents.7. All human material should be considered potentially infectious. All products

derived from human blood are prepared exclusively from the blood of donorstested individually and shown to be free from HBsAg and antibodies to HCVand HIV. The testing methods applied were FDA-approved or cleared incompliance with the European Directve 98/79/EC, Annex II, List A. However, asno testing is absolute, the material should be treated just as carefully as patientspecimen.

8. Do not use multicalibrator over the expiration date/stability period.9. MSDS is available upon request10. Use clean and dry micropipette tips and glassware to dispense multicalibrator.

NOTES

1. Bacterial contamination of the reconstituted multicalibrator will causereductions in the stability of many components.(Criterion for the stability is;recovery within ±5% of initial value)

2. Bilirubin and Acid phosphatase are light sensitive.3. Creatinine Kinase is sensitive to light and temperature.4. For alkaline phosphate assay, allow the reconstituted multicalibrator to stand

for one hour at room temperature5. Inaccurate reconstitution and errors in assay technique can cause improper

calibration.6. Excessive turbidity may indicate microbial contamination in which case the vial

should not be used.

BIBLIOGRAPHY

1. Directive 2000/54/EC. Official Journal of the European Communities No. L262October 17,2000

2. ISO/DIS 17511. In vitro Diagnostic Medical Devises origin-MetrologicalTraceability of values assigned to calibrators and control materials. Geneva,International Organization for standardization 2000.

3. SRM for clinical Chemistry of the National Institute of Standards & Technology(NIST), Gaithersburg, Maryland, USA.

MULTICALIBRATOR5 x 3 mL

11610001

ADL/V.01/APR 2012

98

INTENDED USE

This reagent is intended for the preparation of calibration curve for Ferritin.

REAGENT COMPOSITION

Ferritin calibrator is prepared by diluting human plasma with phosphate bufferedsaline.


The sealed vials are stable up to expiry date at 2-80C, protected from light.


Ferritin calibrator is available in liquid form. Concentration will be mentioned onthe vials. Opened vials are stable for 4 weeks when stored at 2-80C.


Prepare the following calibrator dilution using NaCI as diluent. Multiply theconcentration of the ferritin calibrator by the corresponding factors stated in the tablebelow to obtain the ferritin concentration of each dilution.


Calib.(µL) - 10 10 25 50 100

Saline(µL) 100 150 70 75 50 -

Dil. Factor 0 0.0625 0.125 0.25 0.5 1.0

PRECAUTION


WARNING



BIBLIOGRAPHY

1. Cook, J.D., Lipschitz, D.A. Laughton, M.B.B., Miles, E.M. & Finch, C.A. Serumferritin as a measure of iron stores in normal subjects. A.M.J.clin Nutr 27:680-1974

2. Walters, G.O. Miller, F.M & Wormwood, M.Serum ferritn concentration and ironstores in normal subjects J. Clin pathol 26 770-1973

FERRITIN CALIBRATOR1 x 1 mL

11620003

ADL/V.01/APR 2012

99

INTENDED USE

The reagent is intended for the calibration of photometric systems with HbA1cDirect kit.

REAGENT COMPOSITION

Four levels Hemoglobin A1c calibrator set.

Lyophilized hemolysate prepared from human erythrocytes. Stabilizers to maintainhemoglobin in the reduced state for the accurate calibration of the HbA1cprocedure.

Values are LOT specific please refer to table with lot specific assay data.


The sealed vials are stable up to the expiry date stated on the label, when storedat 2-8 0C.

PREPARATION & STABILITY OF THE CALIBRATOR

1. Open the vial carefully, avoiding any loss of the lyophilized material.2. Add exactly 500 µl of deionized water (inaccurate reconstitution of the

calibrators and error in assay technique can cause erroneous results).3. Close the vial carefully and gently mix for 10 minutes, or until all material has

dissolved, avoiding the formation of foam. Do not use shaker.4. Reconstituted calibrators should be assayed in the same manner as blood

specimens including the hemolysate procedure (please refer to HbA1c Directpack insert. Code No: 11806001

4.1 Dispense 1 mL hemolysis Reagent (R3) into a tube.4.2 Add 20 µL of calibrators and mix. Allow to stand for 5 minutes.Note: The reconstituted calibrators are stable for 30 days at 2-80C, if protected fromlight and heat. Do not freeze.

PRECAUTION

1. To avoid contamination, use clean laboratory wares.2. Close the vials immediately after use. Protect from light and heat. Improper

handing and /or storage of the calibrators can affect results.

WARNING


BIBLIOGRAPHY

1. Nathan, D.M. clin. Chem.29 pp 466-469 (1983) Engbeak, F.et al. Clin.Chem. 35 pp 93-97 (1989)

2. America Diabets Association : Clinical Practice recommendations (positionstatement). Diabetes care 24 (suppl.1): S33-S55, (2001)

3. Tietz, N.W. Textbook of Clinical chemistry, W.B Saunders Company, p.794- 7795 (1999)

HbA1c DIRECT MULTICALIBRATOR4 x 0.5 mL

11604001

ADL/V.01/APR 2012

100

INTENDED USE

This reagent is intended for the preparation of calibration curve for RF estimation.

REAGENT COMPOSITION

RF calibrator prepared by diluting a high level of RF human plasma with saline. Solutioncontains 0.05 g% Sodium azide as preservative.


The sealed vials are stable up to expiry date at 2-80C, if protected from light.


Calibrator is stable liquid and the concentration will be mentioned on the vial. Generatea calibration curve by diluting the calibrator as follows:


Prepare the following calibrator dilution using NaCI as diluent. Multiply theconcentration of the RF calibrator by the corresponding factors stated in the tablebelow to obtain the RF concentration of each dilution.

Dilution 1 2 3 4 5

Calib.(µL) 20 25 50 75 100

Saline(µL) 140 75 50 25 0

Dil. Factor 0.125 0.25 0.5 0.75 1

PRECAUTION


WARNING



BIBLIOGRAPHY

Frederick Wlofe et al. Arthritis and Rheumatism 1991: 34: 951-960

RF CALIBRATOR1 x 1 mL

11617001

101

INTENDED USE

HbA1c control is an assayed quality control intended for monitoring the precision oflaboratory testing procedures.

CHARACTERISTICS

This product is prepared from human blood. The control is provided in liquid form.To obtain consistent vial-to-vial assay values, the control requires proper storage andhandling as described.


Unopened vial is stable until expiry date stated on the vial label at 2-8oC. Once controlis opened, all the analytes will be stable for 7 days when stored tightly capped at 2-8oC.

EXPECTED VALUES

The expected values mentioned in the label are specific for this lot of product.Determinations were performed under strictly standardized conditions on automatedanalyzers using Agappe reagents.

PROCEDURE

To determine HbA1c a heamolysate must be prepared for each test procedure

1.Dispense 500 µL hemolysis reagent into a tube.

2.Add 10 µL of HbA1c control and mix.

3.Allow to stand for 5 minutes or until complete lysis is evident.

Follow the same procedure instructed for Sample and Calibrator


1. For in vitro diagnostic use only2. Do not pipette by Mouth3. If the control comes in contact with eye or skin, wash off immediately with large

amount of water, and seek medical attention immediately.4. Do not use containers and other materials in the package for any purpose other

than those described herein.5. When discarding the controls dispose of them according to local regulations6. Attend to the normal precautions required for handling all laboratory reagents.7. All human material should be considered as potentially infectious. All products

derived from human blood are prepared exclusively from the blood of donorstested individually and shown to be free from HBsAg and antibodies to HCV andHIV, The testing methods applied were FDA-approved or cleared in compliance withEuropean Directive 98/79/EC, Annex II, List A. However, as no testing with absolutecertainly, the material should be treated just as carefully as patient specimen.

8. Do not use HbA1c Control over the expiration date/stability period. Use clean anddry micropipette tips and glassware to dispense HbA1c Control.

HbA1c CONTROL LEVEL 1 & 22 x 0.5 mL

11625002

102

INTENDED USE

Immunology control is an assayed quality control serum intended for monitoring theprecision of laboratory testing procedures.

CHARACTERISTICS

This product is prepared from human serum with added serum proteins. It is astabilized liquid product manufactured under rigid quality control standards. To obtainconsistent vial-to-vial assay values, the control requires proper storage and handlingas described.


Unopened vial is stable until expiry date stated on the vial label at 2-8oC. Once opened,this product will be stable for 30 days when stored tightly capped at 2-8oC.

EXPECTED VALUES

The expected values mentioned in the value sheet are specific for this lot of product.Determinations were performed under strictly standardized conditions on automatedanalyzers using Agappe reagents.


1. For in vitro diagnostic use only2. Do not pipette by mouth3. If the control comes in contact with eye or skin, wash off immediately with large

amount of water, and seek medical attention immediately.4. Do not use containers and other materials in the package for any purpose other

than those described here in.5. When discarding the reagents dispose of them according to local regulations6. Attend to the normal precautions required for handling all laboratory reagents.7. All human material should be considered as potentially infectious. All products

derived from human blood are prepared exclusively from the blood of donorstested individually and shown to be free from HBsAg and antibodies to HCV andHIV, The testing methods applied were FDA-approved or cleared in compliance withEuropean Directive 98/79/EC, Annex II, List A. However, as no testing can rule outthe presence with absolute certainty, the material should be treated just ascarefully as patient specimen.

8. Do not use immunology control over the expiration date/stability period.9. Use clean and dry micropipette tips and glassware to dispense immunology

control.

PROTEIN CONTROL ( IMMUNOLOGY CONTROL )1 x 1 mL

11614005

103

INTENDED USE

Qualicheck Norm & Path is an assayed human serum used for quality controlchecking of clinical chemistry assays suitable for manual procedure and automatedanalyzers.

CHARACTERISTICS

Qualicheck Normal or Pathologic is lyophilized control sera based on human serum.The values given for routine methods, referred to hereafter as specified values, aredetermined under routine conditions according to the method indicated usingdifferent manual or automated assay instructions.

PREPARATION

Open the bottle very carefully, avoiding any loss of the lyophilized material andadd exactly 5 mL dist. water. Close bottle carefully and let stand for 30 minutes.Dissolve contents completely by swirl gently, avoiding the formation of foam. DONOT SHAKE.

STABILITY

The expiry date for the lyophilized control serum is given on the pack

The components of the reconstituted control serum are stable for:-

At least 8 hours at +250C

At least 3 days at +40C

At least 1 month at - 200C when frozen once.

Bilirubin in the reconstituted control serum when stored in the dark is stable for:-

At least 2 hours at + 250C

At least 6 hours at 40C

At least 2 weeks at – 200C when frozen once.

NOTES

This product has been prepared exclusively from the blood of donors testedindividually by FDA approved methods and shown to be free from HbsAg andantibodies to HIV 1 & 2.

As the risk of infection cannot be excluded with certainty, the product must behandled just as the patient specimen.

QUALICHECK NORM & PATH2 x 5 mL

11601001

ADL/V.01/APR 2012

104

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofAlpha1- Acid Glycoprotein in human serum





Alpha-1-acid Glycoprotein is an acute-phase serum protein that is produced by theliver in response to inflammation and infection. AGP is useful in monitoring tumorrecurrence. Levels are also helpful in differentiating acute phase responses (elevatedlevels) from estrogen effects (normal or depressed levels). In addition, it is anexcellent protein in assessing in vivo hemolysis.

PRINCIPLE

The reagents containing polyclonal goat antihuman AGP when mixed with the serumsample containing AGP cause changes in absorbance, due to the development ofturbidity, which is directly proportional to the concentration of Alpha 1- AcidGlycoprotein in the sample.

REAGENT COMPOSITION




Sodium azide(0.95 g/L)



Polyclonal goat anti-human Alpha 1- Acid Glycoprotein

(variable)



The reagents are stable until expiry date when kept at 2-80C.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light.

NORMAL RANGE



Male : 50 - 130 mg/dL


SAMPLE


CALIBRATION

Agappe Protein calibrator (11614002) is recommended for calibration of this assay.


Dilute the high concentrated calibrator 1/10 using saline and use this dilutedcalibrator for the preparation of calibration curve.

Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the AGP calibrator by the corresponding factor stated in the tablebelow to obtain the AGP concentration of each dilution.


1/10 dil.cal.(µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control

It is recommended to use Agappe protein control(11614003) to verify theperformance of the assay. Each laboratory has to establish its own internal qualitycontrol scheme and procedure for corrective action, if controls do not recover within the acceptable tolerance.

ALPHA 1-ACID GLYCOPROTEIN1 x 25mL /1 x4 mL

12005054


Measuring Range:- 4- 300 mg/dl.

If the concentration is greater than linearity (300 mg/dL), dilute the diluted sample(1/10) with normal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 4.66 1.14 2.45

Inter - Run 2.55

Interference:-

No interference for


Na- citrate upto 1000 mg/dL




BIBLIOGRAPHY

1. Schmid,K. in FW Putman,Editor, The plasma protein,Vol 1,second edition,Academic Press, New York 2975, ppt 184-228

2. Johnson, A.M. et al., J. Clin. Invest., 48 (1969)22933. Dati,F.et al.,Lab.Med. 13 (1989)87

ADL/V.01/APR 2012

Parameters Screen

Test AGP

No

Full Name AGP

Standard No. 6

Reaction Type Endpoint

Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 300 µL

R2 30 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank

Linearity Range 4 300

Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit

Determination Coeff. 0

QC Screen

Control 1

Control 2

ALPHA 1-ACID GLYCOPROTEIN

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofcomplement C3 in human serum.





complement C3 is the central point of the classic and alternative complementpathway.

Complement testing help to diagnose the cause of recurrent microbial infections,angioedema, or inflammation. It may be used to help diagnose and to monitor theactivity of acute or chronic autoimmune diseases such as Systemic LupusErythematosus (SLE).

Decreased levels of C3 are significant in autoimmune disease, immune infectionswith pyrogenic bacteria, bacteremia, neonatal respiratory distress syndrome andcongenital deficiencies.C3 behaves as an acute phase protein hence increased levelsmay found in acute inflammatory reactions.

PRINCIPLE


REAGENT COMPOSITION

C3 R1 1 x 35 mL




C3 R2 1 x 6 mL






PRECAUTION


NORMAL RANGE



Serum 75-135 mg/dL

SAMPLE

Use fresh serum. Dilute sample/control to 1/10 with saline. If the test cannot becarried out on the same day, the serum may be stored at 2-80 C for 48 hours.

CALIBRATION



Dilute the calibrator to 1/10 using normal saline and use this diluted calibrator forthe preparation of the calibration curve. Prepare the following calibrator dilutionusing NaCl as diluent. Multiply the concentration of the C3 calibrator by thecorresponding factors stated in the table below to obtain the C3 concentration of eachdilution.


1/10 Cali. (µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control




If the concentration is greater than linearity (400 mg/dL), dilute the diluted sample(1/10) with normal saline and repeat the assay. Multiply the result with dilution factor.

Prozone Effect : >1000mg/dL

Precision in CV% :

Low Medium High

Intra - Run 2.82 3.43 3.28

Inter - Run 3.71 2.56

Interference:- No interference for


Na-citrate upto 1000 mg/dL




BIBLIOGRAPHY

1. Dati, F. et al., Lab. Med.13, 87 (1989)2. Muller-Eberhard, H.H., Ann. Rev.Biochem.44, 697(1975)3. Lachmann, P.J., Hobart, M.J. and Ashton, W.P. (1973)

C31 x 35mL /1 x6mL

12005051

ADL/V.01/APR 2012

Parameters Screen

Test C3

No

Full Name C3

Standard No. 6


Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 200 µL

R2 30 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

C3

ADL/V.01/APR 2012

INTENDED USE






C4 is a constituent of C3 convertase & C5 convertase.

Decreased levels are found in hereditary angioneurotic odema, immune complexdisease and congenital deficiencies.

PRINCIPLE


REAGENT COMPOSITION

C4 R1 1 x 35 mL

Phosphate buffered saline (pH7.43)



C4 R2 1 x 6 mL






PRECAUTION


NORMAL RANGE




SAMPLE

Use fresh serum. Dilute the sample/control to 1/10 with saline. If the test cannot becarried out on the same day, the serum may be stored at 2-80 C for 48 hours.

CALIBRATION



Dilute the high concentration calibrator to 1/10 with normal saline and use this dilutedcalibrator for the preparation of calibration curve. Prepare the following calibratordilution using NaCl as diluent. Multiply the concentration of the C4 calibrator by thecorresponding factors stated in the table below to obtain the C4 concentration of eachdilution.


1/10 dil.Cali. (µL) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


C41 x35 mL/1x6mL

12005052

ADL/V.01/APR 2012



If the concentration is greater than linearity (80 mg/dL), dilute the diluted sample (1/10) with normal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 4.54 2.18 3.96

Inter - Run 4.17 3.08

Interference:-

No interference for


Na-citrate upto 1000 mg/dL


Turbidity upto 5%



BIBLIOGRAPHY

1. Dati, F. et al., Lab. Med.13, 87 (1989)2. Muller-Eberhard, H.H., Ann. Rev.Biochem.44, 697(1975) Lachmann, P.J., Hobart,

M.J. and Ashton, W.P. (1973)

Parameters Screen

Test C4

No

Full Name C4

Standard No. 6


Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 200 µL

R2 30 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

C4

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofApolipoprotein A1 in serum or plasma.



-Multipoint calibration.


Apo A1 is the main protein component of HDL. Apo A1 activates lecithin cholesterolacyltransferase which catalyses the esterification of cholesterol this can then betransported to the liver, metabolized and excreted. People with atheroscleroticvascular changes frequently exhibit decreased levels of Apo A1. Even if theconcentrations of apolipoprotein B are normal, a decreased ApoA1 level may be arisk factor for atherosclerosis. Decreased levels of ApoA1 also occur indyslipoproteinemias, acute hepatic cirrhosis and insulin treated patients.

PRINCIPLE

Anti-human Apo A1 antisera when mixed with human serum containing Apo A1,react to cause an absorbance change , which is measured by immunoturbidometricprinciple. The change in the absorbance can be interpolated in a calibration curveprepared with different known concentrations of calibrator.

REAGENT COMPOSITION

Apo A1 R1 1 x 35 mL

Phosphate buffered saline(pH 7.43)


Detergent 0.1%


Apo A1 R2 1 x 6 mL


Polyclonal goat anti-human Apo-A1(Variable)



The sealed reagents are stable up to expiry date stated on the label, when storedat 2 - 8oC.

PRECAUTION

To avoid contamination, use clean laboratory wares; use clean dry disposablepipette tips for dispensing, close reagent bottle immediately after use . Avoid directexposure of reagent to light.

REFERENCE RANGE

It is recommended that , each laboratory establish its own

reference values. The following values may be used as guidelines.

Men : 107 - 177 mg/dL

Women : 107 - 205 mg/dL

SAMPLE

Use fresh serum. Dilute sample/control to 1 /10 with saline

CALIBRATION

Agappe ApoA1&B calibrator (11613002) is recommended for calibration of thisassay.


Reconstitute the Apo A1 calibrator with 1 mL of distilled water. The reconstitutedcalibrator is stable for 7 days at 2-8oC.

Dilute the high concentrated calibrator 1/10 using saline and use this dilutedcalibrator for the preparation of calibration curve.

Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the Apo A1 calibrator by the corresponding factors stated in the tablebelow to obtain the Apo A1 concentration of each dilution.


1/10dilCali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control

It is recommended to use Agappe control to verify the performance of the assay.Each laboratory has to establish its own internal quality control scheme andprocedure for corrective action, if controls do not recover with in the acceptabletolerance.



If the concentration is greater than 300mg/dl dilute the diluted sample (1/10) withnormal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 3.05 1.12 1.48

Inter - Run 1.63



Bilirubin 20mg /dL

BIBLIOGRAPHY

1. Tillett. W. S.et al: serological reactions in pneumonia with a non-protein samaticfraction of pneumococcus. J.Exp.Med..52,561(1930)

2. Ziegenhagen G, Drahovshy D. klinishe Bedeutung des Creaktiven protein. Medklin 1983;78:45-50.

3. Rifal. N.Tracy.R.P.Ridker, P.M.clinical efficacy of an Automated high sensitivityC-Reactive protein Assay, Clin. chem. 45:12.

APO A11 x 35/1x6mL

12005030

ADL/V.01/APR 2012

Parameters Screen

Test APO A1

No

Full Name Apo A1

Standard No. 6


Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 225 µL

R2 37 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

APO A1

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofApo B in serum.

-Turbidometric Immuno assay


-Multi point calibration


Apo B is the main protein component of LDL. It is necessary for the reaction withLDL receptors in the liver and on cell walls and thus involved in transportingcholesterol, from the liver to the vessel cells.

Elevated levels of Apo-B are frequently found in atherosclerotic vascular changesand are a risk factor for atherosclerosis.

PRINCIPLE

The reagents containing polyclonal goat antihuman Apo-B antibodies when mixedwith the serum sample containing Apo-B cause changes in absorbance due tothe development of turbidity, which is directly proportional to the concentrationof Apo-B in the sample.

REAGENT COMPOSITION

Apo B R1 1 x 35 mL


Polyethyleneglycol 60 g/L

Detergent 0.1%

Sodium azide. 0.95 g/L

Apo B R2 1 x 6 mL

Polyclonal goat anti-human Apo-B(Variable)


Phosphate buffer pH 7.43


The sealed reagents are stable upto the expiry date mentioned on the label whenstored at 2-8oC.

PRECAUTION

To avoid contamination, use clean laboratory wares, such as pipette tips and testtubes. Close reagent bottle immediately after use. Avoid direct exposure of reagentto light

REFERENCE RANGE

It is recommended that, each laboratory establishes its own reference values. Thefollowing values may be used as reference.

Men : 60 - 138 mg/dL

Women : 52 - 129 mg/dL

SAMPLE

Use fresh serum . Dilute sample/ control to 1/10 in saline

CALIBRATION



Reconstitute the Apo B calibrator with 1 mL of distilled water. The reconstitutedcalibrator is stable for 7 days at 2-8oC. Dilute the high concentrated calibrator 1/10using saline and use this diluted calibrator for the preparation of calibration curve.

Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the Apo B calibrator by the corresponding factor stated in the tablebelow to obtain the Apo B concentration of each dilution.


1/10Dil.cal.(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

APO B 1 x 35mL/1 x 6 mL

12602001



If the concentration is greater than 300 mg/dL, dilute the diluted sample (1/10) withnormal saline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 5.24 1.16 0.91

Inter - Run 1.02

Interference:-No interference for



Bilirubin 20mg /dL

BIBLIOGRAPHY

1. Tillett. W.S. et al: Serological reactions in Pneumonia a non-protein somaticfraction of pneumococcus. J. Exp.Med..52,561(1930)

2. Ziegenhagen G, Drahovshy D. klinishe Bedeutung des Creaktiven protein.Med klin 1983;78:45-50.

3. Rifal. N.Tracy.R.P.Ridker, P.M.clinical efficacy of anAutomated High sensitivity C-Reactive protein Assay Clin chem. 45:12.

ADL/V.01/APR 2012

Parameters Screen

Test APO B

No

Full Name Apo B

Standard No. 6


Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 225 µL

R2 37 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

APO B

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofCeruloplasmin in serum.





Ceruloplasmin is a copper oxidase enzyme, important in regulating the ionic stateof iron and other metallic ions. Levels are decreased in hepatolenticulardegeneration or Wilson's disease and Menke's kinky hair syndrome. Levels areelevated by the acute phase response and particularly by estrogens.

PRINCIPLE

The reagents containing polyclonal goat antihuman ceruloplasmin when mixed withthe serum sample containing ceruloplasmin cause changes in absorbance due tothe development of turbidity, which is directly proportional to the concentrationof Ceruloplasmin in the sample.

REAGENT COMPOSITION







Polyclonal goat antihuman Ceruloplasmin



The sealed reagents are stable upto the expiry date mentioned on the label whenstored at 2-8oC. Do not freeze.

PRECAUTION


REFERENCE RANGE

It is recommended that, each laboratory should establishes its own reference values.The following value may be used as a reference.

serum : 22 - 61 mg/dL

SAMPLE

Fresh serum.

CALIBRATION



Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the Ceruloplasmin calibrator by the corresponding factors stated inthe table below to obtain the Ceruloplasmin concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


CERULOPLASMIN 1 x 35 mL/1 x6 mL

12005032



If the concentration is greater than 100 mg/dl , dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 6.31 2.07 2.20

Inter - Run 3.91

Interference:-

No interference for


Na - citrate upto 1000 mg/dL



Triglycerides upto 2500 mg/dL

BIBLIOGRAPHY

Poulik , M.D and Kleiss M.L in The plasma proteins”, F.W. Putman, editor,Vol. 2second edition, Academic press, New York, PP 52-108

ADL/V.01/APR 2012

Parameters Screen

Test Ceruloplasmin

No

Full Name Ceruloplasmin

Standard No. 6


Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 225 µL

R2 37 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CERULOPLASMIN

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofPrealbumin in human serum




Prealbumin is a serum and cerebrospinal fluid carrier of the thyroid hormonethyroxine (T4) and retinol. So the more accurate name for prealbumin istransthyretin .

Prealbumin is formed in the liver, it is a measure of hepatocyte function. Decreasedand increased levels of serum prealbumin are associated with liver disease and areaffected by the existence and degree of liver diseases.

The half life of prealbumin is approximately 2 days, making prealbumin a moretimely and sensitive indicator of protein status.

PRINCIPLE

Antibodies to prealbumin are combined with prealbumin in the patient’s serum,forming immune complexes.The immune complexes cause an increase in the lightscattering with the concentration of prealbumin in the serum. The light scatteringis measured by reading turbidity at 340 nm and 700 nm.

REAGENT COMPOSITION

Prealbumin R1(Buffer Solution) 1 x 25 mL


Prealbumin R2 (Anti Serum Solution) 1 x 4 mL

Anti- human prealbumin anti serum(Variable)



PRECAUTION


NORMAL RANGE





SAMPLE


CALIBRATION



Dilute the high concentrated calibrator to 1/10 using saline and use this dilutedcalibrator for the preparation of calibration curve. Prepare the following calibratordilutions using NaCl as diluent. Multiply the concentration of the Prealbumincalibrator by the corresponding factor stated in the table below to obtain thePrealbumin concentration of each dilution.


Cali.1/10 dil(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control






Precision in CV%:-

PREALBUMIN 1 x25mL /1 x 4 mL

12005055

Low Medium High

Intra - Run 5.30 3.04 3.96

Inter - Run 6.22 4.71 4.1

Interference:-

No interference for



Bilirubin 20mg /dL

BIBLIOGRAPHY

1. Japanese Journal of Clinical Medicine,53(enlarged) , 186 – 188 (1995)2. Journal of Clinical Experimental Medicine , 149 (5) , 277 – 279 (1989)

ADL/V.01/APR 2012

Parameters Screen

Test Pre Alb

No

Full Name Pre Alb

Standard No. 6


Primary WL 340

Sec. WL 670

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 300 µL

R2 30 µL

Sample 15 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

PREALBUMIN

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofTransferrin in human serum





Transferrin(TF) is synthesised in Liver. The main role of Transferrin is to deliveriron from absorption centres to all tissues. Increased serum Transferrin is associatedwith iron deficiency anemia, malignant tumor, polycythemia rubra. Increased levelsof serum transferrin are observed in patients with aplastic anemia, hemolyticanemia and hepatitis. Absence of Transferrin in the body creates a rare geneticdisorder known as atransferrinemia; a condition characterized by anemia andhemosiderosis in the heart and liver that leads to many complications includingheart failure.

PRINCIPLE

Antibodies to human TF are combined with transferrin in the patients serum,forming immune complexes. The immune complexes cause an increase in lightscattering which correlate with the concentration of TF in the serum. The lightscattering is measured by reading turbidity at 340 nm.

REAGENT COMPOSITION

Transferrin R1(Buffer solution) 1 x 25 mL


Transferrin R2(Anti serum solution) 1 x 4 mL

Anti- human TF anti serum (Variable)


The reagents are stable until expiry date when kept at 2-80 C .

PRECAUTION


NORMAL RANGE



Male : 190 - 300 mg/dL


SAMPLE

Use fresh serum. Dilute serum/control to 1/10 with saline.

CALIBRATION



Dilute the high concentrated calibrator 1/10 with normal saline and use this dilutedcalibrator for the preparation of calibration curve.

Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the Transferrin calibrator by the corresponding factors stated in thetable below to obtain the Transferrin concentration of each dilution.


1/10dilcali.(µl) - 10 10 25 50 100

Saline (µL) 100 150 70 75 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1

Quality Control


TRANSFERRIN 1 x 25mL /1 x 4mL

12005056





Precision in CV%:-

Low Medium High

Intra - Run 4.78 1.30 0.78

Inter - Run 4.64 2.46 0.9


No interference for



Lipemia upto 5 % of intralipid

BIBLIOGRAPHY

1. K. Bergstorm, et al.: J.Clin Lab Invest., 40, 637-640 (1980)2. H. Maikus, et al.: Clinica Chemica Acta, 88, 523-530(1978)

ADL/V.01/APR 2012

Parameters Screen

Test Trans

No

Full Name TRF

Standard No. 6


Primary WL 670

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.01

R1 300 µL

R2 30 µL

Sample 8.5 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

TRANSFERRIN

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofIgM antibodies in serum.





IgM is important in early response to infections . The measurement of IgM isimportant for typing immuno deficiencies and myelomas. IgM plays an importantrole in the humoral defense of the body. Serum levels may be increased in all kindof acute infections. Elevated levels in cord serum suggest clinical infection in thenew born.

PRINCIPLE

Antibodies to IgM are combined with IgM in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgM in the serum. The light scattering ismeasured by reading turbidity at 340 nm and 700 nm.

REAGENT COMPOSITION

IgM R1 (Buffer solution) 1 x 30 mL


IgM R2 (Antiserum solution) 1 x 10 mL

Anti-human IgM antiserum


The sealed reagents are stable upto the expiry date mentioned on the label whenstored at 2-8 0C. Do not freeze.

PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure ofreagent to light.


REFERENCE RANGE

It is recommended that each laboratory establish its own reference values. Thefollowing values may be used as reference.


SAMPLE

Fresh Serum.

CALIBRATION



Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the IgM calibrator by the corresponding factor stated in the tablebelow to obtain the IgM concentration of each

Dilution 1 2 3 4 5 6Cal. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


IgM 1 x 30/1 x 10 mL

12005037



If the concentration is greater than 500 mg/dL, dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 1.43 1.59 0.64

Inter - Run 3.17 3.65 2.49

Interference:-

No interference for



Bilirubin 20mg /dL

Turbidity 5 %

BIBLIOGRAPHY

1. Otani,H. :Medical Technology,14, 965(1986)2. Rinsho Kensa Guide 1992, 269-274(1992) Published by BUNKODO CO., LTD3. Kanai’s manual of Clinical laboratory Medicine, the 30 th edition,868-

873(1993)published by KANEHARA & CO.,LTD

ADL/V.01/APR 2012

Parameters Screen

Test IgM

No

Full Name IgM

Standard No. 6

Reaction Type EP

Primary WL 340

Sec. WL 670

Direction Increase

Reaction Time -1 22

Incubation Time 18

Unit mg/dL

Precision 0.1

R1 210 µL

R2 70 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low *

High *

* Data entered by the operator

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

IgM

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofIgG antibodies in serum.


-High linearity of 2700 mg/dL



IgG is a predominant serum immunoglobulin . The measurement of IgG is importantfor typing immunodeficiencies and myelomas. Increased levels are found in chronicinfections and chronic inflammation. IgG is the only immunoglobulin which crossesthe placenta and is therefore of special importance in infants defense againstinfection.

PRINCIPLE

Antibodies to IgG are combined with IgG in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgG in the serum. The light scattering ismeasured by reading turbidity at 700 nm.

REAGENT COMPOSITION

IgG R1 (Buffer solution) 1 x 15 mL


IgG R2 (Antiserum solution) 1 x 15 mL

Anti-human IgG antiserum


The sealed reagents are stable upto the expiry date mentioned on the label whenstored at 2-8 0C.

PRECAUTION


REFERENCE RANGE

It is recommended that , each laboratory should establish its own reference values.The following value may be used as a reference.

serum : 870 - 1700 mg/dL

SAMPLE


CALIBRATION



Dilute the high concentration calibrator to 1/10 using normal saline and use thisdiluted calibrator for the preparation of calibration curve.

Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the IgG calibrator by the corresponding factor stated in the tablebelow to obtain the IgG concentration of each dilution.


1/10dil.Cal(µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


IgG 1 x 15mL/1 x 15 mL

12005036



If the concentration is greater than 2700 mg/dL, dilute the diluted sample (1/10) withnormal saline and repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 2.5 3.25 4.18

Inter - Run 4.08 1.83

Interference:-

No interference for



Bilirubin 20mg/dL

BIBLIOGRAPHY

1. Otani,H. :Medical Technology,14, 965(1986)2. Rinsho Kensa Guide 1992, 269-274(1992) Published by BUNKODO CO., LTD3. Kanai’s manual of Clinical laboratory Medicine, the 30 th edition,868-


ADL/V.01/APR 2012

Parameters Screen

Test IgG

No

Full Name IgG

Standard No. 6

Reaction Type EP

Primary WL 670

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 3

Unit mg/dL

Precision 0.1

R1 180 µL

R2 180 µL

Sample 4 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low *

High *


Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

IgG

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofIgA antibodies, in serum.





The measurement of IgA is important for typing immunodeficiencies and myelomas.Further more it plays a role in acute and chronic infections as first line of defence.Increased levels may be found in acute infectious hepatitis, chronic aggressivehepatitis, cryptogenic cirrhosis, active alcoholic cirrhosis, chronic infections,rheumatoid arthritis, mixed connective tissue diseases etc.

PRINCIPLE

Antibodies to IgA are combined with IgA in the patient’s serum, forming immunecomplexes. The immune complexes cause an increase in light scattering whichcorrelate with the concentration of IgA in the serum. The light scattering ismeasured by reading turbidity at 700 nm.

REAGENT COMPOSITION

IgA R1(Buffer solution) 1 x 30 mL


IgA R2(Antiserum solution) 1 x 10 mL

Anti-human IgA antiserum



PRECAUTION


REFERENCE RANGE

It is recommended that , each laboratory should establish its own reference values.The following values may be used as reference.

Serum : 110 - 410 mg/dL

SAMPLE

Use fresh Serum

CALIBRATION



Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the IgA calibrator by the corresponding factor stated in the tablebelow to obtain the IgA concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


IgA 1 x 30mL/1 x 10mL

12005035





Precision in CV%:-

Low Medium High

Intra - Run 0.84 0.94 1.20

Inter - Run 1.70 1.41 1.07

Interference:-

No interference for



Bilirubin 20mg /dL

BIBLIOGRAPHY

1. K.Bergstorm, et al.: Scand. J. Clin. Lab. Invest., 637(1980)2. Rinsho Kensa Guide 1992, 269-274(1992) Published by BUNKODO CO., LTD3. Kanai’s manual of Clinical laboratory Medicine, the 30 th edition,868-


ADL/V.01/APR 2012

Parameters Screen

Test IgA

No

Full Name IgA

Standard No. 6

Reaction Type EP

Primary WL 670

Sec. WL

Direction Increasing

Reaction Time -1 22

Incubation Time 3

Unit mg/dL

Precision 0.1

R1 210 µL

R2 70 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low *

High *


Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

IgA

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofIgE in human serum and plasma samples.


-Linearity up to 1000 IU/mL



IgE is an immunoglobulin with a molecular weight of approximately 190 KD. It isnormally present in the blood in trace amounts. IgE, like all immunoglobulins, isproduced by plasma cells in response to antigenic stimuli. However abnormal IgE levelsoften results in the development of clinically important Type 1 allergic reactions suchas asthma, hay fever, dermatitis and food allergies. Elevated levels are also seen in casesof parasitic infections, pulmonary aspergillosis, Wiskott-Aldrich Syndrome, hepatitisand myeloma.

The measurement of IgE in human serum is thus considered to be useful in thediagnosis, treatment, assessment of disease progression, or post-operativeprognosis for such conditions.

PRINCIPLE

When an antigen-antibody reaction occurs between IgE and anti-IgE antibody whichhas been coated on latex particles, agglutination results. This agglutination isdetected as an absorbance change, with the magnitude of the change beingproportional to the quantity of IgE in the sample. The actual concentration is thendetermined by interpolation from a calibration curve prepared from calibrators ofknown concentration.

REAGENT COMPOSITION

IgE R1 1 x 22mL


IgE R2 1 x 6.5 mL

Latex suspension

0.125 % w/v suspension of latex

particles sensitized with anti IgE

antibodies (mouse)



PRECAUTION


REFERENCE RANGE



Normal value in Serum/ Plasma : up to 358 IU/mL

SAMPLE

Use Serum / Plasma

CALIBRATION

Agappe IgE calibrator is recommended for calibration of this assay.


Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the IgE calibrator by the corresponding factors stated in the tablebelow to obtain the IgE concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


IgE 1 x 22mL/1 x 6.5mL

12005045


Measuring Range:- 25 –1000 IU/mL.

If the concentration is greater than 1000 IU/mL , dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.

Prozone Effect:- >50000IU/mL

Precision in CV%:-

Low Medium High

Intra - Run 1.18 0.36 0.82

Inter - Run 4.56 1.88 1.21

Interference:-

No interference for



Bilirubin 20 mg /dL

BIBLIOGRAPHY

1. Human neutrophils synthesize IL-8 in an IgE-mediated activation Monteseirin Jet al. J Leukoc Biol. 76: 692-700, 2004

2. Myeloperoxidase release after allergen-specific conjunctival challengeMonteseirin J et al. J Asthma. 41: 639-643, 2004

3. IgE-dependent release of myeloperoxidase by neutrophils from allergic patientsMonteseirin J et al. Clin Exp Allergy. 31 (6): 889-891, Jun 2001

ADL/V.01/APR 2012

Parameters Screen

Test IgE

No

Full Name IgE

Standard No. 6

Reaction Type FT

Primary WL 578

Sec. WL 670

Direction Increase

Reaction Time 1 9

Incubation Time 3

Unit IU/mL

Precision 0.1

R1 200µL

R2 50 µL

Sample 3 µL

R1 Blank 0

Mixed Rgt.Blank 0


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day)

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

IgE

ADL/V.01/APR 2012

INTENDED USE : The reagent is intended for in vitro quantitative determination ofFerritin in serum

-Latex Enhanced Immunoturbidimetry

-High Linearity of 1000 ng/mL



Ferritin is an iron-containing protein. It is mainly found in liver and spleen, whereits function is to eliminate and store iron in the body. It is also found in smallamounts in human serum. The serum levels tend to increase due to hepatitis andmalignant tumors. The measurement of ferritin is useful in diagnosis, treatment,assessment of disease progression and post operative prognosis of abnormal ironmetabolism and iron deficiency anaemia.

PRINCIPLE

Latex particles coated with anti-ferritin antibody are agglutinated when mixed withsamples containing Ferritin. The agglutination causes an absorbance change whichdepends on the Ferritin concentration in the sample, this can be interpolated usinga calibration curve prepared from calibrators of different concentrations.

REAGENT COMPOSITION


Glycine buffer


Suspension of latex particle bound to anti-ferritin antibodies



PRECAUTION

To avoid contamination, use clean laboratory wares, use clean dry disposablepipette tips for dispensing, close reagent bottle immediately after use. Avoid directexposure of reagent to light.

NORMAL RANGE

It is recommended that, each laboratory should establish its own reference values.The following value may be used as a guide line.

Male : 30 - 220 ng/mL

Females : 20 - 110 ng/mL

SAMPLE

Fresh Serum

CALIBRATION

Agappe Ferritin calibrator (11620002) is recommended for calibration of this assay.


Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the Ferritin calibrator by the corresponding factor stated in the tablebelow to obtain the Ferritin concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


FERRITIN1 x 20mL/1 x 11mL

12005044


Measuring Range:- 10 –1000 ng/mL.

If the concentration is greater than 1000 ng/mL , dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.

Prozone Effect:- >30000ng/mL

Precision in CV%:-

Low High

Intra - Run 10 7

Inter - Run 12 10

Interference:-

No interference for

Hemoglobin 500mg/dL


Bilirubin 30mg /dL

Rheumatoid factor up to 560 IU/mL

BIBLIOGRAPHY

1. Cook, J.D., Lipschitz,D.A., Laughton, M.B.B., Miles, E.M. & Finch, C.A: Serumferritin as a measure of iron stores in normal subjects. Am.J.clin.Nutr. 27: 680,1974.

2. Walters,G.O.,Miller, F.M & Wormwood, M. : Serum ferritin concentration onand iron stores in normal subjects. J.clin.pathol. 26: 770-, 1973.

ADL/V.01/APR 2012

Parameters Screen

Test Ferittin

No

Full Name Ferittin

Standard No. 5

Reaction Type EP

Primary WL 578

Sec. WL

Direction Increase

Reaction Time 0 30

Incubation Time 3

Unit ng/L

Precision 0.1

R1 180 µL

R2 90 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

FERRITIN

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofCystatin C in human serum.


-Linear of 10 mg/L


Cystatin C is a low molecular weight (13 Da) cytoplasmic protein, functioning as aninhibitor of various cystein protease in the blood stream. Cystatin C has a stableproduction rate and is removed from the blood circulation by glomerular filtration.In healthy individuals Cystatin C is completely reabsorbed and degraded in the tubulesbut in subject with renal disorders its level in blood may be raised as high as 2 to 5times the normal values. Cystatin C is superior to serum creatinine as a marker ofglomerular filtration Rate.

PRINCIPLE

Cystatin C in the test sample binds to the specific polyclonal rabbit anti-Cystatin Cantibody, which has been adsorbed to latex particle and agglutinates. Theagglutination is detected as absorbance change at 546 nm. The magnitude of changeis proportional to the quantity of Cystatin C in the sample and its concentration isdetermined by interpolation from a calibration curve prepared from calibrators ofknown concentration.

REAGENT COMPOSITION


Tris buffer 1.2% (100 mM)

pH 8.5+0.3

Cystatin C R2 1 x 5.5 mL

Polystyrene latex particle coated with polyclonal anti Cystatin C antibody (rabbit)

Reagents required but not provided:

Cystatin C Calibrator (Product code -11623003)



PRECAUTIONS:


NORMAL RANGE



Individuals up to 50 years : 0.55 – 1.15 mg/L

Individuals above 50 years : 0.63 – 1.44 mg/L

SAMPLE

Required sample material is human serum or EDTA/ Heparinized plasma. It isrecommended to analyze the sample as fresh as possible.

CALIBRATION

Agappe Cystatin C calibrator is recommended for calibration of this assay.

Dilution of calibrator for calibration curve:

Calibration Curve (range between 0-10 mg/L). Prepare the following calibratordilution using NaCI as diluent. Multiply the concentration of the Cystatin C calibratorby the corresponding factor stated in the table below to obtain the Cystatin Cconcentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 200 190 70 60 50 -

Dil. factor 0 0.05 0.125 0.25 0.5 1.0


Measuring Range:- 0.1 - 10 mg/L.

If the concentration is greater than 10 mg/L, dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.


Precision :

MEAN INTRA RUN INTER RUN nVALUE CV(%) CV(%)







CYSTATIN-C 1 x 23mL/1 x 5.5 mL

12005043

Interference:-

No interference for


Intrafat 1400 mg/dL

Bilirubin 25 mg /dL

BIBLIOGRAPHY

Filler G,.Bokenkamp A, Hofmann W, Le Bricon T, Martnez – Bru C, Grubb ADharnidharka VR, Kwon Stevens G.

ADL/V.01/APR 2012

Parameters Screen

Test Cys - C

No

Full Name Cystatin -C

Standard No. 6

Reaction Type EP

Primary WL 546

Sec. WL 670

Direction Increase

Reaction Time 0 18

Incubation Time 3

Unit mg/L

Precision 0.1

R1 200 µL

R2 40 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank

Linearity Range 0.1 10

Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CYSTATIN-C

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofHbA1c in human blood.



-Direct result in % HbA1c from analyzer

-No total Hb determination required


HbA1c is a glycated form of haemoglobin formed by the attachment of glucoseresidues in the blood to the hemoglobin molecules. In the diabetic populationwhere blood glucose levels are abnormally elevated the level of HbA1c alsoincreases. The level of HbA1c is proportional to the level of glucose in the bloodand has been widely accepted as an indicator of the mean blood glucoseconcentration in the preceeding 6-8 weeks. It is therefore a long-term indicator ofdiabetic control. For routine use HbA1c levels should be monitored every 3-4months. However in gestational diabetes and after a change in therapy it may beuseful to measure HbA1c more frequently at 2-4 week intervals.

PRINCIPLE

This method utilizes the interaction of antigen and antibody to directly determinethe HbA1c in whole blood. Total heomoglobin and HbA1c have the samenonspecific absorption rate to latex particle. When mouse antihuman HbA1cmonoclonal antibodies are added (R2), latex HbA1c – mouse antihuman HbA1cantibody complex is formed. Agglutination occurs when goat anti mouse IgGpolyclonal antibody interacts with the monoclonal antibody. The amount ofagglutination is proportional to the amount of HbA1c absorbed onto the surfaceof latex particles. The amount of agglutination is measured as absorbance, whichis used to calculate HbA1c % from a calibration curve.

REAGENT COMPOSITION

HbA1c R1 1 x 30 mL

Latex 0.13%(w/v)

Glycine buffer 20 mmol.L

HbA1c R2A 1 x 9.5 mL


HbA1c R2B 1 x 0.5 mL

Mouse anti-human HbA1c 10 mmol/L

Monoclonal antibody

Goat anti-mouse IgG 0.05 mg/dL

(Polyclonal)

Stabilizers 0.08 mg/dL

HbA1c R3 2x 63 mL

Haemolysis Reagent


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C, protected from light.

LINEARITY

The reagent is linear up to 16% (NGSP)

NORMAL RANGE



According NGSP :

<6% for non-diabetic

< 7% for glycemic control of person with diabetes

According to IFCC:

< 4.2% for a nion-diabetic

< 5.3% for glycemic control of a person with diabetes.



Reagent R2 is prepared by pouring the entire contents of the R2B vial into the R2Avial. Mix gently. This working reagent is stable for 30 days at 2-80C.

CALIBRATION

Agappe HbA1C Direct Multicalibrator ( 11604001) is recommended for calibrationof this assay. Reconstitute the calibrator with 0.5 mL distilled water and it will bestable up to 30 days at 2-80C. Do not freeze.

Quality Control

It is recommended to use Agappe HbA1C Control level 1&2 (11604003) to verifythe performance of the assay. Each laboratory has to establish its own internal qualitycontrol scheme and procedure for corrective action, if controls do not recover within the acceptable tolerance.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposablepipette tips for dispensing. Close reagent and standard bottles immediately afteruse.


SAMPLE

Whole blood, collected with EDTA

To determine HbA1c a heamolysate must be prepared for each sample

1. Dispense 1mL hemolysis reagent into a tube.2. Add 20 µL of well-mixed whole blood and mix.3. Allow to stand for 5 minutes or until complete lysis is evident.

Follow the same procedure with calibrators and controls.INTERFERENCES

No interference up to :

Ascorbic acid 50 mg/dL

Bilirubin 50 mg/dL

Triglycerides 2000 mg/dL

Carbamylated Hb 7.5 mmol/L

Acetylated Hb 5.0 mml/L


BIBLIOGRAPHY

1. Nathan, D.M., Clin, Chem. 29, pp.466-469 (1983)2. Engbeak, F., et al. Clin chem.35 pp. 93-97 (1989)3. American Diabetes Association : Clinical practice recommendations (position

statement). Diabetes care 24 (suppl.1) S33-S55, (2001).4. Tietz, N.W. Textbook of Clinical Chemistry, W.B. Saunders Company,

p.794 -7795 (1999).

HbA1c DIRECT 1 x 30/1 x 9.5/1 x 0.5/2x63mL

12005029

ADL/V.01/APR 2012

Parameters Screen

Test HbA1C D

No

Full Name HbA1C Direct

Standard No. 5


Primary WL 630

Sec. WL

Direction Increase

Reaction Time -1 19

Incubation Time 18

Unit %

Precision 0.01

R1 210 µL

R2 75 µL

Sample 6 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low *

High *


Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

HbA1c DIRECT

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofAnti Streptolysin – O (ASO).


-Linear up to 800 IU/mL.


ß-hemolytic streptococcus bacteria especially group A, C and G, produce an exotoxinknown as Streptolysin-O. People infected with this bacterium produce an antibodyknown as Anti Streptolysin-O (ASO). Measuring the levels of ASO is effective fordiagnosing, judging the progress of medical treatment and assessing the recoveryfrom diseases like rheumatic fever, acute glomerulonephritis and tonsillitis.

PRINCIPLE

When an antigen-antibody reaction occurs between ASO in the sample andstreptolysin-O which has been sensitized to latex particles, agglutination occurs. Thisagglutination results in change of absorbance and is proportional to the quantityof ASO in the sample. The actual concentration is determined by interpolation froma calibration curve prepared from calibrators of known concentration.

REAGENT COMPOSITION

ASO R 1 1 x 20 mL


ASO R2 1 x 11 mL

ASO Latex suspension particles coated with Streptolysin - O



PRECAUTION

To avoid contamination use clean laboratory wares. Use clean dry disposable pipettetips for dispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light.

NORMAL RANGE



Serum

Adults : 200 IU/mL

children < 5 years : 100 IU/mL

CALIBRATION

Agappe ASO Calibrator ( Product code-11615002) is recommended for calibrationof this assay.

Quality Control


SAMPLE

Fresh serum / plasma

ASO 1 x 20/1 x 11 mL

12005040


Measuring Range:- 20–800 IU/mL.

If the concentration is greater than 800 IU/mL, dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Medium High

Intra - Run 5.0 3.0

Inter - Run 8 5

INTERFERENCES

Do not interfere for



Intrafat up to 5000 mg/dL

BIBLIOGRAPHY

1. Galuin, J.P. et al.: Particle enhanced photometric immune assay system, Clin.Lab Assays (pap .Annu.clin.Lab.Assays Conf.)

2. Singer J.M. et al. The latex fixation test Application to the serologic diagnosisor rheumatoid arthritis , Amer J.Med 21, 888(1956)

ADL/V.01/APR 2012

Parameters Screen

Test ASO

No

Full Name ASO

Standard No. 2


Primary WL 578

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time 10

Unit IU/mL

Precision 0.01

R1 180µL

R2 90µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen w

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

ASO

ADL/V.01/APR 2012

INTENDED USE : This reagent is intended for in vitro quantitative determination ofC-reactive protein in human serum or plasma by immunoturbidimetry.




CRP (C – reactive Protein) is a cytokine - induced, acute phase protein thatincreases in concentration as a result of inflammation. CRP levels in the body hasbeen used as a marker or indicator of infections and inflammation. The assay ofCRP is more sensitive than the erythrocyte sedimentation rate (ESR) and leukocytecount. The CRP levels rise and return to reference ranges more rapidly after thedisease has subsided.

PRINCIPLE

This is a latex enhanced turbidimetric immuno assay. CRP in the samples binds tospecific anti-CRP antibodies, which have been adsorbed to latex particles andagglutinates. The agglutination is proportional to the quantity of CRP in the sample.The actual concentration is then determined by interpolation from a calibrationcurve prepared from calibrators of known concentrations.

REAGENT COMPOSITION

CRP R1 1 x 20 mL

Glycine buffer

CRP R2 1 x 8 mL

Latex suspension coated with anti-CRP antibodies. (rabbit polyclonal antibody)



PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure ofreagent to light.


NORMAL RANGE

It is recommended that each laboratory should establish its own reference values.The following value may be used as a guide line.

Serum up to 6 mg/L

SAMPLE

Fresh serum (Do not use hemolized or lipemic serum)

CALIBRATION

Agappe CRP Calibrator ( 11616002) is recommended for calibration of this assay.


Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the CRP calibrator by the corresponding factor stated in the tablebelow to obtain the CRP concentration of each dilution.


Cali. (µL) - 10 10 20 50 100

Saline (µL) 100 150 70 60 50 -

Dil. factor 0 0.0625 0.125 0.25 0.5 1.0

Quality Control


CRP 1 x 20/1 x8 mL

12005041


Measuring Range:- 1 –200 mg/L.

If the concentration is greater than 200mg/L, dilute the sample with normal saline andrepeat the assay. Multiply the result with dilution factor.

Prozone Effect:- >1000mg/L

Precision in CV%:-

Low Medium High

Intra - Run 7.0 5.0 3.0

Inter - Run 10 8 5

Interference:-

No interference for

Hemoglobin 500mg/dL

Intrafat 500 mg/dL

Bilirubin 30mg /dL

RF up to 500 IU/mL

BIBLIOGRAPHY

1. Tillett.W.S..et al: Serological reactions in pneumonia with a non protein somaticfraction of pneumococcus.J.Exp.Med..52,561(1930).

2. Zeigenhagen G,Drahovshy D.Klinishe Bedeutung des C-reaktiven protein.Medklin1983;78:45-50.

3. Rifal.N.Tracy.R.P.Ridker,P.M.Clinical efficacy of an Automated High sensitivity CReactive protein Assay..45-12.

ADL/V.01/APR 2012

Parameters Screen

Test CRP LEIT

No

Full Name CRP LEIT

Standard No. 6


Primary WL 578

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time 10

Unit mg/L

Precision 0.01

R1 210 µL

R2 70 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen w

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CRP

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofRheumatoid factor in Serum.


-Linear upto 135 IU/mL


Rheumatoid Factor (RF) is an auto antibody against human IgG commonly seen insera, particularly in patients with rheumatoid arthritis. The measurement of RF valueis useful in evaluating the diagnosis, effects of therapy and prognosis of RA, systemiclupus erythematosus, Chronic hepatopathy etc.

PRINCIPLE

When a sample containing rheumatoid factor is added to denatured human IgGwhich has been sensitizied to latex particles, antigen-antibody reaction occursleading to agglutination. This agglutination leads to an absorbance change whichis measured at (550 to 660nm). The change in absorbance is proportional toagglutination and the actual concentration is determined by interpolation from acalibration curve prepared from known value calibrators.

REAGENT COMPOSITION

RF R 1 1 x 20 mL

Glycine Buffer Solution

RF R2 1 x 8 mL

Latex suspension coated with denatured human IgG



PRECAUTION

To avoid contamination, use clean laboratory wares. Avoid direct exposure ofreagent to light & heat.


NORMAL RANGE



Serum up to 18 IU/mL

SAMPLE

Fresh serum (Do not use haemolized or lipemic serum)

CALIBRATION

Agappe RF Calibrator ( Product code-11617002)) is recommended for calibrationof this assay.


Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the RF calibrator by the corresponding factor stated in the tablebelow to obtain the RF concentration of each dilution.


Cali. (µL) - 10 20 50 75 100

Saline (µL) 100 70 60 50 25 -

Dil. factor 0 0.125 0.25 0.5 0.75 1.0

Quality Control


RF 1x 20/1 x 8 mL

12005048


Measuring Range:- 10 - 135 IU/mL.

If the concentration is greater than 135 IU/mL, dilute the sample with normal salineand repeat the assay .Multiply the result with dilution factor.


Precision in CV%:-

Low High

Intra - Run 5.0 1.0

Inter - Run 8.0 5.0

Interference:-

No interference for

Hemoglobin 500mg/dL

Intrafat 5000mg/dL

Bilirubin 20mg /dL

BIBLIOGRAPHY

Frederick Wolfe et al-Arthritis and Rheumatism 1991 : 34:951-960

ADL/V.01/APR 2012

Parameters Screen

Test RF

No

Full Name RF

Standard No. 6


Primary WL 578

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time 10

Unit IU/dL

Precision 0.01

R1 210 µL

R2 70 µL

Sample 6 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

RF

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofLipoprotein (a) in serum.

-Multipoint calibration with fixed time mode

-Linearity up to 80 mg/dL


Lp(a) is a low density lipoprotein like particle containing apoliprotein B-100disulphide-linked to one large glycoprotein called apoliprotein(a). Many investigatorshave confirmed that a high lipoprotein(a) concentration represents an indicator ofrisk for cardio vascular diseases, especially when, the serum LDL-cholesterol or apoB are elevated. The quantification of Lp (a) in serum or plasma is important foridentification of individuals at risk for developing artherosclerosis.

PRINCIPLE

Latex particles coated with anti-human Lp(a) are agglutinated when mixed withsamples containing Lp(a). The agglutination causes an absorbance changedependent upon the Lp(a) contents of the patient sample, that can be interpolatedin a calibration curve prepared with different calibrators of different Lp(a) contents.

REAGENT COMPOSITION


Buffer solution (pH 8.3)

LIPOPROTEIN (a) R2 1 x 5.5 mL

Lipoprotein (a) latex

Reagents required but not provided:

Lp(a) Calibrator ( Product code-11619002)



PRECAUTION



NORMAL RANGE



Serum up to 30 mg/dL

SAMPLE

Fresh Serum sample (free of haemolysis)

CALIBRATION

Agappe Lp(a) Calibrator ( 11619002) is recommended for calibration of this assay.

Calibration curve : Prepare dilutions of the Lp(a) calibrator using 9 g/L saline asdiluent:

Dilution 1 2 3 4 5

Cali. (µL) - 25 50 75 100

Saline (µL) 100 75 50 25 -

Dil. factor 0 0.25 0.5 0.75 1.0

Multiply the Lp(a) calibrator concentration by the corresponding dilution factorindicated in the table to obtain the Lp(a) concentration of the different (diluted)calibrators.


1. Measurement Range: 12-80mg/dLIf the concentration is greater than linearity (80mg/dL), dilute the sample andrepeat the assay. Multiply the result with dilution factor.

2. Prozone effect: No prozone effect was detected up to 225 mg/dL.INTERFERENCES

Bilirubin : up to 427 mmol/L no interference

Hemoglobin : up to 10 g/L no interference

Lipids : up to 5 g/L no interference

BIBLIOGRAPHY

1. Gaubalz JW, et al. J.Biol Chem 1983; 258 45832 -45892. Berg K A Acta Pathol Microbiol Scand 1963:59:369-3823. Scanu AM , et al. J.Clin invest 1990;85: 1709 -1715

Lp (a) 1 x 23/1 x 5.5 mL

12005046

ADL/V.01/APR 2012

Parameters Screen

Test LP(a) LEIT

No

Full Name LP(a) LEIT

Standard No. 6

Reaction Type EP

Primary WL 670

Sec. WL

Direction Increase

Reaction Time 0 18

Incubation Time 3

Unit mg/dL

Precision 0.1

R1 200 µL

R2 40 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low *

High *


Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

Lp (a)

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofmicroalbumin in human urine

-Turbidometric Immunoassay




Albumin is normally found in the blood. When the kidneys are working properly,albumin will not be present in the urine. However, when the kidneys are damaged,small amounts of albumin leak into the urine. This condition is calledmicroalbuminuria.

Microalbuminuria is most often caused by kidney damage from diabetes. However,many other conditions can lead to kidney damage, such as high blood pressure,heart failure, cirrhosis, or systemic lupus erythematosus (SLE). If early kidneydamage is not treated, larger amounts of albumin and protein may leak into theurine. This condition is called macroalbuminuria or proteinuria, this can lead tochronic kidney disease.

PRINCIPLE

The reagents containing polyclonal goat antihuman microalbumin when mixed withthe urine sample containing microalbumin cause changes in absorbance, due tothe development of turbidity, which is directly proportional to the concentrationof microalbumin in the sample.

REAGENT COMPOSITION


Saline (9 g/L)

Accelerator


Microalbumin R2 1 x 5.5 mL

Phosphate buffered saline

Polyclonal goat anti-human albumin (variable)




PRECAUTION

To avoid contamination, use clean laboratory materials. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use. Avoid directexposure of reagent to light. Do not Freeze.

NORMAL RANGE



Urine : 0 -25 mg/L (IFCC)

SAMPLE

Use fresh Urine.

CALIBRATION

Agappe Microalbumin Calibrator ( Product code-11618002) is recommended forcalibration of this assay.


Prepare the following calibrator dilutions using NaCl as diluent. Multiply theconcentration of the microalbumin calibrator by the corresponding factor stated inthe table below to obtain the microalbumin concentration of each dilution.


Calib.(µL) - 10 10 25 50 100

Saline(µL) 100 150 70 75 50 -

Dil. Factor(µL) 0 0.0625 0.125 0.25 0.5 1.0

MICROALBUMIN 1 x 23/1 x 5.5 mL

12005047


Measuring Range:- 4 - 395 mg/L

If the concentration is greater than linearity (395 mg/L), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.


Precision in CV%:-

Low Medium High

Intra - Run 2.28 1.8 3.04

Inter - Run 2.93 0.66 0.53

Interference:-

No interference for

Hemoglobin 1000mg/dl

Bilirubin 10mg/dL

BIBLIOGRAPHY

1. Mount,J., J. Clin. Pathology, 22, 12 (1986)2. Schmidtz, A., et al., diabetic Medicine, 5 , 126 (1988)

ADL/V.01/APR 2012

Parameters Screen

Test MicroAlb

No

Full Name Micro Albumin

Standard No. 6

Reaction Type Fixed time

Primary WL 340

Sec. WL

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/L

Precision 0.01

R1 200 µL

R2 40 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

MICROALBUMIN

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofC-reactive protein (CRP) in serum.

-Latex Enhanced Immuno Turbidimetric assay

-Sensitivity of 0.13 mg/L

-Linearity up to 10 mg/L


CRP is an acute phase protein produced by liver. It’s level will rise in response toinflammations and infections.

Inflammation of arteries is a risk factor for cardiovascular disease. It is linked to anincreased risk of heart disease, heart attack, stroke and peripheral arterial disease.CRP ultra/HS CRP is the strongest predictor of cardiac risk. The value more than 10mg/L cannot be considered for cardiac risk factors.

Routinely available CRP methods are to determine infections or chronicinflammatory disease where CRP concentration is above 10 mg/L. These methodsare with limited sensitivity hence it cannot precisely measure CRP concentrationsbelow 10 mg/L.

Studies have shown that measuring CRP with improved methodology of highsensitive assay can identify the risk level of CVD in apparently healthy people.Relatively high levels of hs CRP in healthy individuals are predictive of the futurerisk of heart disease even when cholesterol levels are within the acceptable range.

PRINCIPLE

This is a latex-enhanced turbidimetric invitro immuno assay. CRP in the sample bindsto specific anti-CRP antibodies, which had been adsorbed to latex particles andagglutinates. The agglutination is detected as an absorbance change. The magnitudeof the change is proportional to the concentration of CRP in the sample. The actualconcentration is then detected by interpolation from a calibration curve preparedfrom calibrators of known concentration.

REAGENT COMPOSITION


Glycine buffer


Latex suspension coated with anti-CRP antibodies. (Rabbit polyclonal antibody)


The sealed reagents are stable up to the expiry date stated on the label, when storedat 2-80C. Stability in the instrument is at least 4 weeks if contamination is avoided.Do not freeze.

PRECAUTION

To avoid contamination, use clean laboratory wares. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use.


REFERENCE RANGE



Less than 1 mg/L = Low risk for CVD

1.0-2.9 mg/L = Intermediate Risk for CVD

Greater than 3 mg/L = High Risk for CVD

SAMPLE


CALIBRATION

Agappe CRP Ultra calibrator is recommended for calibration of this assay.


Prepare the following calibrator dilution using NaCl as diluent. Multiply theconcentration of the CRP ultra calibrator by the corresponding factors stated in thetable below to obtain the CRP ultra concentration of each dilution.


Calib.(µL) - 10 10 20 50 100

Saline(µL) 100 150 70 60 50 -

Dil. Factor 0 0.0625 0.125 0.25 0.5 1.0

CRP Ultra 1x20 / 1x11 mL

12005042


Measuring Range:- 0.13 – 10mg/L.

Prozone Effect:- >1000mg/L

Precision in CV%:-

Low Medium High

Intra - Run 7 5 3

Inter - Run 10 8 5




Conjugated Bilirubin up to 30 mg/dL

Intra Fat up to 500 mg/dL

Rheumatoid Factor up to 500 IU/mL

BIBLIOGRAPHY

1. Claus DR,Osmand AP, Gewurz H. Radioimmunoassay of human C-reactiveprotein and levels in normal sera. J Lab Clin Med 1976;87: 120-128

2. Wasunna A, Whitelaw A,Gallimore R,Hawkins PN,Pepys MB. C-reactive proteinand bacterial infection in preterm infants. Eur J Pediatr 1990; 149: 424-427

ADL/V.01/APR 2012

CRP Ultra

Parameters Screen

Test CRP

No

Full Name CRP

Standard No. 6


Primary WL 578

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time 10

Unit mg/L

Precision 0.01

R1 180 µL

R2 90 µL

Sample 3 µL

R1 Blank

Mixed Rgt.Blank

Linearity Range 0.13 10

Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low

High

Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

ADL/V.01/APR 2012

SyRe - BS120/200 - Updated

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Albumin in serumor plasma.

-Bromocresol green methodology

-Linear up to 6 g/dL


Albumin which is synthesized in the liver constitutes a major part of the totalproteins in the body, the other part being globulin; they form the major portion ofthe dissolved substances in the plasma. Functions of Albumin includes distributionof extracellular fluid, regulation of osmotic pressure, acts as transport agent for awide variety of substance such as hormones lipids, vitamins etc.

Increased levels are seen in dehydration.

Decreased levels are seen in liver disease (Hepatitis, Cirrhosis), malnutrition, kidneydisorders, increased fluid loss during extensive burn & malabsorption.

PRINCIPLE

The reaction between albumin from serum or plasma and the dye bromocresol-green produces a change in colour that is proportional to the albumin concentration

REAGENT COMPOSITION

ALBUMIN REAGENT 4 x 30 mL

Succinate Buffer (pH 4.20) 75 mmol/L

Bromocresol green 0.14 g/L

Reagents Required But not Provided

Multi Calibrator ( Product Code No - 11610001)

Qualicheck path (Product Code No - 11601002)

Qualicheck norm (Product Code No - 11601003)

STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat room temperature.

LINEARITY

This reagent is linear upto 6 g/dL

If the concentration is greater than linearity (6 g/dL), dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum/plasma – 3.5 – 5.5 g/dL

CALIBRATION

Agappe Multicalibrator( Product Code No - 11610001)is recommended forcalibration .

Quality Control – It is recommended to use Qualicheck Norm (Code No. 11601003)or Qualicheck path (Code No. 11601002) to verify the performance of themeasurement procedure.

Each Laboratory has to establish its own internal quality control scheme andprocedures for corrective action if controls do not recover within the acceptabletolerance.

BIBLIOGRAPHY

1. Doumasa B.T.etal: Clin. Chim Acta 31, 87 pp (1971)

2. Weis, W.A. :Klin. Wochenschr. 43, S.273 (1965)

ALBUMIN4 x 30 mL

12005001

ADL/V.01/APR 2012

Parameters Screen

Test ALB

No

Full Name Albumin

Standard No. 2

Reaction Type End Point

Primary WL 630

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time

Unit g/dL

Precision 0.1

R1 250 µL

R2 0

Sample 3 µL

R1 Blank 0 3000

Mixed Rgt.Blank

Linearity Range 0 6

Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low *

High *

Calibration Screen

Rule Two Point Linear

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

Albumin

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of AlkalinePhosphatase in serum or plasma.

-DGKC – SCE recommended procedure .

-Linear up to 700 U/L


Alkaline phosphatase is widely distributed throughout the body, but clinicallyimportant one for diagnostic reasons are in bone, liver, placenta & intestine.Growing bone is associated with the release of ALP and so in childhood the levelof ALP is around 3 times of that of adult. During pregnancy in 2nd & 3rd trimesterthe enzyme rises considerably due to placenta releasing ALP. It can be used toexamine placental function.

Elevated levels are seen in bone diseases, e.g. Paget’s disease, Rickets, Osteoblasticmetastatic & in obstructive disease of biliary tract.


ALP = Alkaline Phophatase

PRINCIPLE

Kinetic determination of Alkaline Phosphatase(ALP) is based upon the followingreaction.

ALP

Para nitrophenyl phosphate + H2O ------------> p-nitrophenol+Inorganic phosphate


REAGENT COMPOSITION


Diethanolamine Buffer,(pH10.2) 125 mmol/L

Magnesium Chloride 0.625mmol/L


P-Nitrophenyl phosphate 50 mmol /L

REAGENTS REQUIRED BUT NOT PROVIDED

Multi Calibrator (Product Code No - 11610001)





LINEARITY

The reagent is linear, up to 700 U/L

If the concentration is greater than linearity (700 U/L), dilute the sample withnormal saline & repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE

It is recommended that each laboratory establish its own reference values. Thefollowing value may be used as guideline.

Women 64 – 306 U/L

Men 80 – 306 U/L

Children up to 15yrs < 644 U/L

PRECAUTION



SAMPLE

Fresh serum/plasma (free of haemolysis)

CALIBRATION



Each Laboratory has to establish it’s own internal quality control scheme andprocedures for corrective action if controls do not recover within the acceptabletolerance.

BIBLIOGRAPHY

1. Schlebush, H.et al.,Dtsh.med.Wschr.99,765(1974)2. Z.Klin. Chem.,Klin. Biochem.8,658(1980)10,182(1972)

ALKALINE PHOSPHATASE2 x 30 / 2 x 8 mL

12005002

ADL/V.01/APR 2012

Parameters Screen

Test ALP

No

Full Name Alk. Phosphatase

Standard No. 2

Reaction Type Kinetic

Primary WL 405

Sec. WL

Direction Increase

Reaction Time 4 12

Incubation Time 10

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 5 µL

R1 Blank 0 10000

Mixed Rgt.Blank


Linearity Limit 0.2

Substrate Limit 13000

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

ALKALINE PHOSPHATASE

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Amylase in serum,plasma & urine.

-CNPG3 methodology.

-Linear up to 2000 U/L


Amylase occurs in the salivary glands, fallopian tubes & in pancreas.

a -amylase is secreted by the pancreas from where it enters the duodenum, throughthe pancreatic duct. Any obstruction to these ducts causes

a-amylase enzyme to enter the blood stream.

Elevated levels are seen in acute pancreatitis, peptic ulcers, biliary disease, parotitis& other intestinal obstructions.

Decreased levels are seen in chronic pancreatic disorders having pancreatic celldestruction.

PRINCIPLE

Amylase

5CNPG3 -------------> 3 CNP +2CNPG

2+3 Maltotriose + 2 Glucose.

CNP : Chloro-4-nitrophenol

CNP-G2 : 2-chloro -4-nitrophenyl-a maltoside

REAGENT COMPOSITION

ALPHA AMYLASE (S.L) R1 2 x 30 mL

MES Buffer (pH6.0) 50mmol/L

CNPG3

2.27 mmol/L



Activator 900 mmol/L







LINEARITY

The reagent is linear, up to 2000 U/L

If the concentration is greater than linearity (2000 U/L), dilute the sample withnormal saline & repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE


Serum , plasma 25-86 U/L

Urine < 470 U/L

PRECAUTION



This reagent is very sensitive to external contamination, ie Saliva, Sweat etc whichcontains a-amylase. Handle with gloves & keep vial tightly sealed after use.

Discard reagent if it turns cloudy.

SAMPLE

Fresh serum, plasma (free of haemolysis)

Urine (1/3 diluted)

CALIBRATION




BIBLIOGRAPHY

1. Junge, W. et al., Clin. Biochem. 22, 109(1989)2. Hohenwallnern, W., J.Clin. chem.. Clin. Biochem. 27,97(1989)

AMYLASE 2 x 30 mL

12005003

ADL/V.01/APR 2012

Parameters Screen

Test Amy

No

Full Name Amylase

Standard No. 2


Primary WL 405

Sec. WL

Direction Increase

Reaction Time 4 12

Incubation Time

Unit U/L

Precision 0.1

R1 200 µL

R2 0

Sample 5 µL

R1 Blank 0 2000

Mixed Rgt.Blank


Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

AMYLASE

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Bilirubin in serumor plasma .



Bilirubin is formed by the breakdown of RBC’s in the spleen, liver & bone marrow.Small amount of bilirubin circulates in the plasma loosely bound to albumin, whichis not water soluble. This is referred to as indirect or unconjugated bilirubin. In theliver bilirubin is conjugated with glucuronic acid, which forms a soluble compound.This is referred to a direct bilirubin.


PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. DirectBilirubin reacts with diazotized sulfanic acid to form azobilirubin.

REAGENT COMPOSITION



Hydrochloric acid 165 mmol/L

Preservatives and stabilizers






STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. Activator should be stored at 2 - 80C

LINEARITY



NORMAL RANGE



Direct Bilirubin - up to 0.4 mg/dL

PRECAUTION



SAMPLE


CALIBRATION




BIBLIOGRAPHY

1. Water M., Gerard H.: MICROCHEM JM 15, 231(1980)2. Annino J. S.: C.C. Principles and procedure,19603. A.A. A.C.C.: Clin Chem 8 : 405,196

BILIRUBIN DIRECT 4 x 30 / 2 x 8 mL

12005004

ADL/V.01/APR 2012

Parameters Screen

Test Bili D

No

Full Name Bilirubin Direct

Standard No. 2


Primary WL 546

Sec. WL 630

Direction Increase

Reaction Time -1 20

Incubation Time 10

Unit mg/dL

Precision 0.1

R1 300 µL

R2 35 µL

Sample 15 µL

R1 Blank 0 1000

Mixed Rgt.Blank 0 1200


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

BILIRUBIN DIRECT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Bilirubin in serumor plasma

-Modified TAB method



Bilirubin is formed by the breakdown of RBCs in the spleen, liver & bone marrow.Small amount of bilirubin circulates in the plasma loosely bound to albumin, whichis not water soluble. This is referred to as indirect or unconjugated bilirubin. In theliver bilirubin is conjugated with glucuronic acid, which forms a soluble compound.This is referred to as direct bilirubin.


PRINCIPLE

Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. TotalBilirubin reacts with diazotized sulfanilic acid in the presence of TAB to formazobilirubin.

REAGENT COMPOSITION



TAB 9 mmol/L







STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat RT. Activator should be stored at 2 - 80C

LINEARITY



NORMAL RANGE




PRECAUTION



SAMPLE


CALIBRATION




BIBLIOGRAPHY

1. Walter M., Gerard H.: MICROCHEM JM 15, 231.(1980)2. Annino J. S.: C.C. Principles and procedure,19603. A.A. A.C.C.: Clin Chem 8 : 405,196

BILIRUBIN TOTAL (TAB) 4 x 35 / 2 x 10 mL

12005005

ADL/V.01/APR 2012

Parameters Screen

Test Bili T

No

Full Name Bilirubin Total

Standard No. 2


Primary WL 546

Sec. WL 630

Direction Increase

Reaction Time -1 20

Incubation Time 10

Unit mg/dL

Precision 0.1

R1 300 µL

R2 35 µL

Sample 15 µL

R1 Blank 0 1000



Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

BILIRUBIN TOTAL

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Calcium in serum,plasma & urine.

-Modified Arsenazo III method




Increased levels are found in hyperthyroidism, malignant tumors, acute &osteoporosis, adrenal insufficiency.

Decreased levels are found in hypoparathyrodism, osteomalacia, rickets, renalfailure & tetanus.

PRINCIPLE

At a neutral pH the Ca2+ form with Arsenazo III a complex, the color intensity ofwhich is directly proportional to the concentration of calcium in the sample.

REAGENT COMPOSITION

CALCIUM ARSENAZO REAGENT 2 x 35 mL

MES, pH 6.50 1000 mmol/L

Arsenazo III 200 mmol/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum / plasma : 8.8 – 10.2 mg/dL

Urine : 100-400 mg/24 hrs

SAMPLE


Urine diluted 1/3 with distilled water; adjust to pH 3-4 with HCI (N/10).

Take dilution factor into account for the calculation of the concentration in urine.

CALIBRATION




BIBLIOGRAPHY

Baver, P.J. Anal. Biochem., 110, (1981), 61

CALCIUM (ARSENAZO)2 x 35 mL

12005006

ADL/V.01/APR 2012

Parameters Screen

Test Cal

No

Full Name Calcium A

Standard No. 2


Primary WL 630

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2 0

Sample 3 µ L

R1 Blank 0 15000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CALCIUM (ARSENAZO)

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Chloride in serum,plasma & urine.

-Modified Thiocyanate method

-Linear up to 130 mEq/L


Chloride & bicarbonate are the principle anions (-vely charged) where as sodium &potassium are the principle cations (+vely charged) in the plasma. Chloride ions areinvolved in regulation of water distribution between the tissues by maintainingosmotic pressure & normal cation & anion balance between intra & extra cellularfluids.

Elevated levels are seen in conditions like dehydration & congestive cardiac failure.

Decreased levels are seen in condition such as salt losing nephritis, diabetic acidosis& renal failure.

PRINCIPLE

In an acid medium chloride ions and mercury ( II) thiocynate form thiocynate ions.These ions react with HNO

3 and iron (III) ions and effect a red color. The intensity

of the color is directly proportional to the concentration of chloride ions.

REAGENT COMPOSITION

CHLORIDE REAGENT 2 x 30 mL

Mercuric (II) thiocyanate 2 mmol/L

Nitric acid 29 mmol/L

Ferric Nitrate 20 mmol/L





STORAGE & STABILITY


LINEARITY

This reagent is linear up to 130 mEq/L

If the concentration is greater than linearity (130 mEq/L), dilute the sample withdistilled water and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE


The following values may be used as guideline.

Serum : 97 - 108 mEq/L

Urine : 120 - 240 mEq/L/24 hr

SAMPLE

Serum / plasma (free of haemolysis) / Urine (Dilute sample 1:1 with distilled waterand multiply the result with 2)

CALIBRATION




BIBLIOGRAPHY

Schonfed R.G.Lowellen C.S: Clin Chem.10,533 pp (1964)

CHLORIDE 2 x 30 mL

12005007

ADL/V.01/APR 2012

Parameters Screen

Test Chloride

No

Full Name Chloride

Standard No. 2


Primary WL 510

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time

Unit mEq/L

Precision 0.1

R1 250 µL

R2 0

Sample 3 µL

R1 Blank 0 1000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CHLORIDE

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Cholesterol inserum or plasma.

-CHOD-PAP methodology.

-Linear up to 600 mg/dL.

-Contains LCF (Lipamic Clearing factor) which minimizes rerun.


Cholesterol is the main lipid found in the blood, bile & brain tissues. It is also oneof the most important steroids of the body & is a precursor of many steroidhormones. Two thirds of cholesterol present in the blood is esterified. The livermetabolizes the cholesterol & it is transported in the blood stream by lipoproteins.

Increased levels are found in hypercholesterolemia, hyperlipidemia, hypothyroidism,uncontrolled diabetes, nephritic syndrome & cirrhosis.

Decreased levels are found in malabsorption, malnutrition, hyperthyroidism,anaemia & liver diseases.

PRINCIPLE

Enzymatic determination of total cholesterol according to the following reactions.

CHE

Cholesterol ester +H2O ------------> Cholesterol + fatty acids

CHO

Cholestrol + O2

----------------> 4-Cholesten-3- one + H2O

2

POD

2H2O

2 +Phenol+4-Aminoantipyrine-------------> Red quinone + 4H

2O

CHE : Cholesterol Esterase

CHO : Cholesterol Oxidase

POD : Peroxidase

REAGENT COMPOSITION

CHOLESTEROL (S.L) R1 4 x 35 mL


Phenol 24 mmol/L

Sodium Cholate 0.5 mml/L

Cholesterol Esterase > 180 U/L

Cholesterol Oxidase > 200 U/L


4 – aminoantipyrine 0.5 mmol/L





STORAGE & STABILITY


LINEARITY


If the concentration is greater than linearity ( 600 mg/dL), dilute the sample withnormal saline and repeat the assay. Multiply with dilution factor

NORMAL RANGE



Serum, Plasma : 150 – 220 mg/dL

SAMPLE

Serum, Plasma (free of haemolysis).

CALIBRATION




BIBLIOGRAPHY

Allain C.C. et al., Clin.Chem 20 (1974), 470

CHOLESTEROL 4 x 35 mL

12005008

ADL/V.01/APR 2012

Parameters Screen

Test CHO

No

Full Name Cholesterol

Standard No. 2


Primary WL 510

Sec. WL 630

Direction Increase

Reaction Time 0 24

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2 0

Sample 3 µL

R1 Blank 0 2000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low 150

High 220

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit

Determination Coeff.

QC Screen

Control 1

Control 2

CHOLESTEROL

ADL/V.01/APR 2012

CREATININE 4 x 35 / 2 x 18 mL

12002055

INTENDED USE: This reagent is intended for in vitro quantitative determination ofcreatinine in serum, plasma & urine.


It is formed in muscles from phospho creatinine. It is important form of energy beinga store of high-energy phosphate. Creatinine determinations have one advantage overUrea determination that it is not affected by a high protein diet.

Serum creatinine is more specific & sensitive indicator of renal function. Simultaneousestimations of serum urea & creatinine provides better information. Serum ureanitrogen, creatinine ratio is > 15 in pre renal failure, & < 10 in renal failure.

Decreased levels are found in muscle dystrophy.

PRINCIPLE

Creatinine reacts with picric acid to produce a colored compound, creatinine alkalinepicrate. The change in absorbance is proportional to the creatinine concentration.

REAGENT COMPOSITION

CREATININE DYE REAGENT 4 x 35 mL

Picric Acid 8.73 mmol/L

Surfactant

CREATININE BASE REAGENT 2 x 18 mL


Sodium Phosphate 25 mmol/L

Reagents required but not provided

Multicalibrator ( Product Code No. 11610001)

Qualichek Norm ( Product Code No. 11601003)

Qualichek Path ( Product Code No. 11601002)

STORAGE & STABILITY


This reagent is linear up to (24 mg/dL).


NORMAL RANGE



Serum : Men : 0.7 – 1.4 mg/dL

Female : 0.6 – 1.2 mg/dL

Urine : : 0.80 – 1.80 gm/24 hr

PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / Urine (diluted 1/100 with distilled water)

CALIBRATION

Agappe Multicalibrator (Code No. 11610001) is recommended for calibration of thisassay.



BIBLIOGRAPHY

1. Allen. L.C.: Clin chem.Vol.28 No.3, 1982, 555.2. Haeckel, R. et al. Chlin. Chem. 27/1 179-183 (1981).3. Tanganelli, E. Prencipe, L; Bassi, D. Cambiaghi, S. and Murador,E.: Clin.Chem 287,

1461-1464 (1982)

Parameters Screen

Test CRT

No

Full Name Creatinine

Standard No. 2

Reaction Type Fixed Time

Primary WL 510

Sec. WL

Direction Increase

Reaction Time 4 12

Incubation Time 5

Unit mg/dL

Precision 0.1

R1 200 µL

R2 200 µL

Sample 40 µL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CREATININE

INTENDED USE

This reagent is intended for in vitro Quantitative determination of Creatine Kinasein human serum.

-Optimized IFCC Method.

-Linear up to 1700 U/L.


It is mainly found in all muscle (Cardiac & Skeletal) & brain tissues. It plays animportant role in energy storing mechanism of the tissues. Its iso-enzymes: CK-MBmainly exists in cardic muscle tissues, CK-MM in skeletal muscle tissues & CK-BB inbrain & lungs.

Increased levels are found in myocardical infarction, muscular dystrophy,cerebrovascular-disease, pulmonary infarction, electrical shocks & hypothyrodisim.

Decreased levels are, sometimes seen in early pregnancy.

PRINCIPLE

Kinetic determination of Creatinine Kinase is based on following reactions

CK

Creatinine Phophate + ADP ------------> Creatine + ATP

HK

ATP + D-Glucose --------------------> G-6-p+ADP

G-6-PDH

G-6-P + NADP+ ---------------> D-Gluconate -6-Phosphate + NADPH + H+

CK- Creatinine Kinase

Hk- Hexokinase

G-6-P-D- Glucose-6-phosphate

G-6-PDH- Glucose-6 Phosphate dehydrogenase

REAGENT COMPOSITION

CREATINE KINASE R1 1 x 30 mL

Imidazole (pH 6.7) 125 mmol/L

D-Glucose 25 mmol/L

N-Acetyl-L-Cysteine 25 mmol/L


NADP 2.52 mmol/L

EDTA 2.02 mmol/L

Hexokinase > 6800 U/L

CREATINE KINASE(S.L) R2 1 x 8 mL


ADP 15.2 mmol/L

AMP 25 mmol/L

Diadenosine-5-pentaphosphate 103 µmmol/L

G-6-PDH >88800 U/L





STORAGE & STABILITY


LINEARITY



NORMAL VALUES



Men : < 171 U/L

Women : < 145 U/L

PRECAUTION



SAMPLE

Serum/Plasma (Free of haemolysis)

CREATINE KINASE 1 x 30 / 1 x 8 mL

12005010

CALIBRATION

Agappe Multicalibrator(Product Code No - 11610001)is recommended forcalibration .

Quality Control : It is recommended to use Qualicheck Norm (Code No. 11601003)or Qualicheck path (Code No. 11601002) to verify the performance of themeasurement procedure.

Each Laboratory has to establish its own internal quality control scheme andprocedures for corrective action if controls do not recover within the

acceptable tolerance.

BIBLIOGRAPHY

1. DGKC,J.Clin.chem.clin.Bioch.15,255(1977)2. Di.Witt, Trendelenberg, J.Clin.Chemie,clin. Bioch.20,235(1982)

ADL/V.01/APR 2012

Parameters Screen

Test CK- NAC

No

Full Name CK- NAC

Standard No. 2


Primary WL 340

Sec. WL

Direction Increase

Reaction Time 8 20

Incubation Time 5

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 12.5 µL

R1 Blank 0 10000



Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

CREATINE KINASE

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination creatinine in serumor urine.


-No sample dilution


-MSDS available for analyzers


It is formed in muscles from phosphocreatinine. It is important form of energy beinga store of high energy phosphate. Creatinine determinations have one advantageover urea determination that it is not affected by a high protein diet.

Serum creatinine is more specific & sensitive indicator of renal function.Simultaneous estimations of serum urea & creatinine provides better information.Serum urea nitrogen & creatinine ratio is > 15 in prerenal failure & < 10 in renalfailure. Decreased levels are found in muscle dystrophy.

PRINCIPLE

Creatininase

Creatinine +H2O --------------------------> Creatine

Creatinase

Creatine + H2O ---------------------------> Sarcosine +Urea

Sarcosine Oxidase

Sarcosine + O2+H

2O ----------------------------> Glycine + HCHO + H

2O

2

Peroxidase

2 H2O

2 + 4-AA *1+ TOOS *2 ---------------------> Quinone pigment +4 H

2O

*1 : 4- Aminoantipyrine, * 2N-Ethyl –N-(2-Hydroxy -3-Sulfopropyl)-m-toluidine

CRE concentration can be obtained by measuring quinone pigment photometrically

REAGENT COMPOSITION

CREATININE R1 2 x 35 mL

Creatinase 175000 IU/L

Sacrosine Oxidase 15000 lU/L

TOOS 1.13 mmol

CREATININE R2 2 x 12 mL

Creatininase 75000 IU/L

Peroxidase 4500 units /L

4-AA 0.75 mmol





STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C

LINEARITY


If the concentration is greater than linearity dilute the sample with normal salineand repeat the assay. Multiply the result with dilution factor.

NORMAL VALUES



Serum : Male : 0.6 – 1.1 mg/dL

: Female : 0.5 – 0.8 mg/dL

Urine : Male : 1070–2150 md/dL(24 hrs-accumulated urine)

: Female : 769 – 1200 mg/dL (24 hr accumulated urine)

PRECAUTION



SAMPLE

Fresh Serum/Urine 24 hr

CALIBRATION




ENZYMATIC CREATININE 2 x 35 / 2 x 12 mL

12005012

BIBLIOGRAPHY

Artiss. J.D. Mc Enroe, R.J.Zak B.Clin.Chem, 30 (1984)1389.

ADL/V.01/APR 2012

Parameters Screen

Test Enzy CRT

No

Full Name Enzymatic Creatinine

Standard No. 2


Primary WL 546

Sec. WL 630

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.1

R1 225 µL

R2 75 µL

Sample 6 µL

R1 Blank 0 1500



Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

ENZYMATIC CREATININE

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro Quantitative determination of GAMMA GT inserum.

-Szasz methodology.



GGT activity is elevated in all forms of liver diseases. It is highest in cases ofintrahepatic or post hepatic biliary obstruction. (It may be 5 to 30 times higher thannormal) It is more sensitive than Alkaline Phosphatase , NTP, Leucineaminopeptidase and transaminases in detection of obstructive jaundice, cholangitis,cholecystis neoplasm, it rises earlier than other enzyme and persists longer.

Moderate increase is observed in infectious hepatitis. (2 to 5 times) Increases mayalso be observed in cases of drug intoxication, acute and chronic pancreatitis.

PRINCIPLE

Kinetic determination of Gamma GT according to the following reaction.

Gamma GT

GLUPA-C+Glycylglycine --------------> L-Gamma-Glutamyl-Glycylglycine

+ 5-Amino-2-nitrobenzoic acid.

GLUPA-C: L-Gamma -Glutamyl-3 Carboxy-P-nitroanilide

REAGENT COMPOSITION

Gamma GT R1 1 x 30 ml

Tris buffer pH (8.25) 133 mmol/L

Glycylglycine 138 mmol/L

Gamma GT R2 1 x 8 ml

GLUPA - C 23 mmol/L





STORAGE & STABILITY


LINEARITY


If the concentration is greater than linearity (232 U/L) dilute the sample with normalsaline and repeat the assay. Multiply the result with dilution factor.

NORMAL VALUES



Female : 5-32 U/L

Male : 10-45 U/L

PRECAUTION

To avoid contamination, use clean laboratory wares. .


SAMPLE

Fresh Serum (free of haemolysis)

CALIBRATION




BIBLIOGRAPHY

1. Szasz, G.Clin.Chem., 22, (1976),20512. SCJ.Clin. Lab.Invest 36: 711 (1976)3. Tietz N.W. Text book of Clin.Chem. 678 686 : (1986)

GAMMA GT 1 x 30 / 1 x 8 mL

12005013

ADL/V.01/APR 2012

Parameters Screen

Test GGT

No

Full Name GGT

Standard No. 2

Reaction Type kinetic

Primary WL 405

Sec. WL

Direction Increase

Reaction Time 4 12

Incubation Time 10

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 25 µL

R1 Blank 0 10000



Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

GAMMA GT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Glucose in serum,plasma & CSF

-GOD –PAP methodology



Glucose is a major carbohydrate present in the blood & serves as a primary sourceof energy. It is usually obtained from ingested starch & sugar. The glucoseconcentration is normally maintained at constant level. Excessive glucose is storedas a inactive glycogen mainly in the liver & little in the muscles.



PRINCIPLE


GOD

Glucose+ O2 + 2H

2O --------------------> H

2O

2 + Gluconic acid

POD

2H2O

2+phenol + 4-Ap -------------------> Quinonimine + 4H

2O

GOD – Glucose Oxidase

POD – Peroxidase

REAGENT COMPOSITION

GLUCOSE REAGENT R1 4 x 35 mL

Tris Buffer, (pH 7.40) 92 mmol/L

Phenol 0.3 mmol/L

Glucose Oxidase 15000 U/L

4- Aminophenazone 2.6 mmol/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum : Fasting : 70-110 mg/dL

2 hrs postprandial : 70-140 mg/dL

C S F : 50 -70 mg/dL

PRECAUTION



SAMPLE

Serum / plasma (free of haemolysis) / CSF

CALIBRATION




BIBLIOGRAPHY

1. Trinder, P. Ann Clin Biochem. 6,24 (1979)2. Dingeon, B. Ann.Bio.Clin 33,3 (1975)3. Lott, J. Clin.Chem. 21, 1754 (1975)

GLUCOSE 4 x 35 mL

12602001

ADL/V.01/APR 2012

Parameters Screen

Test Glucose

No

Full Name Glucose

Standard No. 2

Reaction Type EP

Primary WL 510

Sec. WL 630

Direction Increase

Reaction Time 0 38

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2 0

Sample 3 µL

R1 Blank 0 2000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low 70

High 110

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

GLUCOSE

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofHDL Cholesterol in serum


Blood total cholesterol levels have long been known to be related to coronary heartdisease (CHD). In recent years, in addition to total cholesterol, high densitylipoprotein cholesterol (HDL-C) has become an important tool used to assess anindividual risk of developing CHD since a strong negative relationship between HDL-C concentration and the incidence of CHD was reported.

PRINCIPLE

The reaction between cholesterol other than HDL & enzyme for cholesterol assayis suppressed by the electrostatic interaction between polyanions & cationicsubstances. Hydrogen peroxide is formed by the free cholesterol in HDL bycholesterol oxidase. Oxidative condensation of EMSE and 4-AA is caused byhydrogen peroxide in the presence of peroxidise, and the absorbance of theresulting red-purple quinine is measured to obtain the cholesterol value in HDL

Polyanions

Other lipoproteins than HDL------------> Suppress reaction with enzyme

Cationic substances

cholesterol esterase

HDL (cholesterol esters) + H2O ----------------> HDL (free cholesterol) + Free fatty acids

cholesterol oxidase

HDL (free cholesterol) + O2 + H+-------------------> Cholestenone + H

2O

2

Peroxidase

2H2O

2 + 4-AA + EMSE + H

3 + O ---------------> Red-purple quinine + 5H

2O

REAGENT COMPOSITION

HDL - C DIRECT R1 2 x 30 mL

N-Ethyl-N-(3-methylphenyl)-N’succinylethyenediame(EMSE)

HDL - C DIRECT R2 2 x 10 mL

Cholesterol Oxidase

4-Aminoantipyrine (4-AA)





STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 - 80C, protected from light. Do not freeze.

LINEARITY



NORMAL RANGE



Male : 35-80 mg/dL

Female : 42- 88 mg/dL

PRECAUTION

To avoid contamination, use clean laboratory materials. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use.


SAMPLE








(when triglyceride in a sample exceeds 1000 mg/dL, dilute the sample to 1+9 withsaline, repeat the assay and multiply result by 10)

HDL – C DIRECT 2 x 30 / 2 x 10 mL

12005015

CALIBRATION




BIBLIOGRAPHY

1. Williams P et al., High density lipoprotein and coronary risk factor, Lancet.1:72 (1979)

2. Gordon,T.Castelli, W.P.Hjortland, M.C. et al. Am.J.Med 62, 707-714 (1977)3. Rifai, N.and Warnick, G.R., Ed. Laboratory Measurement of Lipids,

Lipoproteins and Apolipoproteins AACC Press. Washington, DC,USA,1994

ADL/V.01/APR 2012

Parameters Screen

Test HDL D

No

Full Name HDL Direct

Standard No. 2


Primary WL 578

Sec. WL 670

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.1

R1 225 µL

R2 75 µL

Sample 3 µL

R1 Blank 0 1000



Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

HDL – C DIRECT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of LactateDehydrogenase in serum or plasma.Based on SCE recommended method.



This enzyme is found in all organ cells, but especially plentiful in cardiac & skeletalmuscle, liver, kidney & RBC. LDH is found in the form of iso-enzymes. Based on theirelectrophoretic mobility with each iso-enzymes being primarily from differentorgans.

Elevated levels are found in myocardial infraction, liver diseases, hemolyticanaemia’s, pernicious anaemia, Leukemia & Pulmonary diseases. Elevations in acuteMI reaches a peack in 48-72 hrs. Delayed elevations, (10-14 days) are useful in thelate diagnosis of the condition.

PRINCIPLE

Kinetic determination of lactate dehydrogenase according to the following reaction.

LDH

Pyruvate + NADH + H+ -------------> L-Lactate +NAD+

REAGENT COMPOSITION

LDH -P R1 1 x 30 mL


Pyruvate 1.6 mmol/L

Sodium Chloride 200 mmol/L

LDH – P R2 1 x 8 mL

NADH 240 mmol/L





STORAGE & STABILITY


LINEARITY


If the concentration is greater than linearity (2400 U/L), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor

NORMAL RANGE



Serum /Plasma : 225-450 U/L

PRECAUTION



SAMPLE

Serum, plasma (free of haemolysis)

CALIBRATION


Quality Control - It is recommended to use Qualicheck Norm (Code No. 11601003)or Qualicheck path (Code No. 11601002) to verify the performance of themeasurement procedure.


BIBLIOGRAPHY

1. Z.Klin chem. Klin Biochem.8,658 (1970), 1, 1820(1972)2. Wei Bhaar, D.et al. Med.Welt 26,387 (1975)

LDH 1 x 30 / 1 x 8 mL

12005016

ADL/V.01/APR 2012

Parameters Screen

Test LDH

No

Full Name LDH

Standard No. 2

Reaction Type kinetic

Primary WL 340

Sec. WL

Direction Decrease

Reaction Time 4 12

Incubation Time 10

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 3 µL

R1 Blank 0 20000



Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

LDH

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of LDL Cholesterolin serum or plasma


Blood total cholesterol levels have long been known to be related to coronary heartdisease (CHD). In recent years, in addition to total cholesterol, low densitylipoprotein cholesterol (LDL-C) has become an important tool used to assess anindividual risk of developing CHD since a strong positive relationship between LDL-C concentration and the incidence of CHD was reported. LDL Cholestrol acts as akey factor in the pathogenesis of atherosclerosis and coronary artery disease.

PRINCIPLE

The LDL-C Direct is a homogeneous assay. When serum is mixed with R1, amphotericsurfactants protect LDL from enzyme reactions. CHE and CO reacts with non-LDLcholesterol, which decomposed to water by catalase. R2 enables the conversationof LDL-C to hydrogen peroxide, which upon oxidative condensation with HDAOS &4AA yields a color complex. By measuring the absorbance of this blue color complexproduced, the LDL-C concentration in the sample can be calculated when comparedwith the absorbance of the LDL-C Calibrator.

REAGENT COMPOSITION

LDL –C DIRECT R1 1 x 30 mL

4-Aminoantipyrin 0.5 mmol/l

CHE

CO 1.2 U/mL

Peroxidase


LDL –C DIRECT R2 1 x 10 mL

N,N-bis(4-sulfobutyl)-m-toluidine

disodium salt (DSBmT) 1.0 mmol/l






STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Desirable < 130 mg/dL

Borderline 130-159 mg/dL

High Risk for CHD > 160 mg/dL.

PRECAUTION

To avoid contamination, use clean laboratory materials. Use clean, dry disposablepipette tips for dispensing. Close reagent bottles immediately after use.


SAMPLE

Fresh serum (free of haemolysis) or EDTA Plasma

INTEREFERING SUBSTANCES




Haemoglobin up to 500 mg/dL


(when triglyceride in a sample exceeds 1000 mg/dL, dilute the sample 1+9 withsaline, repeat the assay and multiply result by 10)

LDL – C DIRECT 1 x 30 / 1 x 10 mL

12005017

CALIBRATION




BIBLIOGRAPHY

1. Crouse J.R et al., Studies of low density lipoprotein molecular weight in humanbeingd with coronary artery disease. J.Lipid Res 26:5666 (1985)

2. Gordon,T.Castelli, W.P.Hjortland, M.C. et al. AM.J.Med 62, 707-714 (1977)Rifai, N.and Warnick, G.R., Ed. Laboratory Measurement of Lipids, Lipoproteinsand Apolipoproteins AACC Press. Washington, DC,USA,1994

ADL/V.01/APR 2012

Parameters Screen

Test LDL D

No

Full Name LDL Direct

Standard No. 2


Primary WL 546

Sec. WL 630

Direction Increase

Reaction Time -1 22

Incubation Time 20

Unit mg/dL

Precision 0.1

R1 225 µL

R2 75 µL

Sample 3 µL

R1 Blank 0 15000



Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

LDL – C DIRECT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Magnesium inserum or plasma.

-Xylidyl Blue with ATCS



Magnesium is the second most abundant intracellular cation of the human bodyafter potassium, being essential in a great number of enzymatic and metabolicprocesses.

It is a co-factor of all the enzymatic reactions that involve ATP and found in themembranes that maintain the electrical excitability of muscular and nervous cells.

A low magnesium level is found in malabsorption syndrome, diureticsaminoglucoside therapy, and hyperparathyroidism or diabetic acidosis.

Elevated concentration of magnesium is found in uremia, chronic renal failure,glomerulo nephritis, Addison’s disease or intensive anti acid therapy.

Clinical diagnosis should not be made on a single test result; it should integrateclinical and other laboratory data.

PRINCIPLE

Magnesium reacts with xylidyl Blue to form a colored compound in alkaline solution.

The intensity of the colour formed is proportional to the magnesium in the sample.

REAGENT COMPOSITION

MAGNESIUM REAGENT 1 x 30 mL

Xylidyl Blue 110 mmol/L

Ethanolamine (pH 11.0) 1 mol/L

GEDTA 60 mmol/L





STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 -80C, protected from light.

LINEARITY



NORMAL RANGE



Serum : 1.8 – 2.6 mg/dL

CSF : 2.1 – 3.3 mg/dL

Urine : 73 - 122 mg/24 h

PRECAUTION

To avoid contamination, use clean laboratory materials. It is recommended to usedisposable tubes. Use clean, dry disposable pipette tips for dispensing. Close reagentimmediately after Use. Avoid direct exposure of reagent to light.

SAMPLE

Serum (free haemolysis) or Heparinized plasma.

(Do not use oxalates or EDTA as anticoagulant)

Urine should be acidified to pH 3-4 with concentrated HCl then dilute sample 1/5with distilled water and multiply the result by 5.

CALIBRATION




BIBLIOGRAPHY

1. Farrel, E.C. Magnesium. Kaplan a et al. Clin chem.. The CV Mosby Co St LouisTorento. Princeton 1984; 1064-69

2. Brutis, C. A. et al. TIETZ textbook of clinical chemistry, 3 rd edition W BSaunders company; 1999, P.1395-1457

3. Young, D. S. Effects of disease on clinical lab tests 4th edition AACC Press, 2001

MAGNESIUM 1 x 30 mL

12005018

ADL/V.01/APR 2012

Parameters Screen

Test Mag

No

Full Name Magnesium

Standard No. 2


Primary WL 546

Sec. WL 0

Direction Increase

Reaction Time 0 20

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2

Sample 3 µL

R1 Blank

Mixed Rgt.Blank

Linearity Range 0 5

Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day)

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

MAGNESIUM

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Phosphorous inserum or plasma.

-Phosphomolybdate methodology



Phosphorous is mainly combined with calcium & is found in bones. It is involved inthe carbohydrate metabolism & is a component of many other substances. Someof its important functions include maintaining of acid-base balance, skeletal muscleformation. It is also required for normal functioning of RBCs & muscles.

Increased levels are found in hypothyroidism, renal failure, bone metastasis & liverdisease.

Decreased levels are found in hyperparathyroidism, osteomalacia & diseaseassociated with Vitamin D deficiency.

PRINCIPLE

Determination of inorganic phosphorous according to the following reaction.

phosphorous

Ammonium molybdate + sulfuricaid ----------------> phosphomolybidiccomplex

REAGENT COMPOSITION

INORGANIC PHOSPHOROUS REAGENT 1 x 30 mL

Sulfuric acid 210 mmol/L

Ammonium molybdate 650 mmol/L





STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat 2 80C.

LINEARITY



NORMAL RANGE



Serum : 2.7 – 4.5 mg/dL

Urine : 400 – 1300 mg/24 hr.



PRECAUTION



SAMPLE

Serum / Plasma (free of haemolysis)

Urine diluted to 1/10 with distilled water

CALIBRATION




BIBLIOGRAPHY

1. Tietz, N., Clinical Guide to Laboratory Tests, W.B. Saunders Company, Philad,1983, 384

2. Henry, R.J.Clin. Chem., Harper & Row Publishers. New Yoork 19743. Thomas, L., Labor and Diagnose, 2 Aufl. Med.Vert. Gem. Marburg 1979 Taussky,

H.H.Schorr, E.,J.Biol. Chem 202, 675 (1953)

PHOSPHOROUS1 x 30 mL

12005019

ADL/V.01/APR 2012

Parameters Screen

Test Phos

No

Full Name Phosphorous

Standard No. 2


Primary WL 340

Sec. WL

Direction Increase

Reaction Time 0 15

Incubation Time

Unit mg/dL

Precision 0.1

R1 250 µL

R2 0

Sample 5 µL

R1 Blank 0 3500

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

PHOSPHOROUS

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGOT in serumor plasma.

-Proven IFCC methodology



It is present in most of the tissues, especially in cardiac muscle, liver cells, skeletalmuscle & kidneys. Injury to these tissues results in the release of the enzyme inblood stream.

Increased levels are found in myocardial infarction. The duration & extent ofincrease is related to the infract. SGOT determination is of considerable value todifferentiate myocardial infraction from other cardiac disorders.

Increased levels are also found in various types of liver disease, skeletal muscletrauma & in renal diseases.


PRINCIPLE


AST

L- Asparate + a - ketoglutarate ------------------->Oxaloacete + L-Glutamate

MDH

Oxaloacetate + NADH + H+ -------------------> L- Malate + NAD+

AST :Aspartate aminotransferase.


REAGENT COMPOSITION

SGOT R1 4 x 35 mL

Tris Buffer (pH7.8) 88 mmol/L


MDH > 900 U/L

LDH > 1500 U/L

SGOT R2 2 x 18 mL

a -ketoglutarate 12 mmol/L

NADH 0.24 mmol/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum up to 46 U/L

PRECAUTION



SAMPLE

Serum/plasma (free of haemolysis)

CALIBRATION




BIBLIOGRAPHY

1. Clin, Chim, Acta 105,147-172 (1980).2. Thefeld,W. et.al.,Dtsh.med.wschr.99,343 (1994)

SGOT4 x 35 / 2 x 18 mL

12005020

ADL/V.01/APR 2012

Parameters Screen

Test SGOT

No

Full Name SGOT

Standard No. 2


Primary WL 340

Sec. WL

Direction Decrease

Reaction Time 4 12

Incubation Time 10

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 10 µL

R1 Blank 0 20000



Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

SGOT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of SGPT in serumor plasma.

-IFCC recommended methodology



It is present in most of the tissues, but mainly found in the liver. Increased levelsare found in hepatitis, cirrhosis, obstructive jaundice & other hepatic disease. SGPTactivity is markedly elevated even before clinical signs of jaundice become apparentin disease associated with hepatic necrosis. Slight elevations are also found inmyocardial infraction.

PRINCIPLE

Kinetic determination of SGPT base on the following reaction.

ALT

L-Alanine + a-ketogutarate -------------------> Pyruvate +L-Glutamate

LDH

Pyruvate +NADH+ H+ ----------------------------> L-Lactate +NAD+



REAGENT COMPOSITION

SGPT R1 4 x 35 mL



LDH > 1500 U/L

SGPT R2 2 x 18 mL

NADH 0.24 mmol/L

a-ketoglutarate 16 mmol/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum up to 49 U/L

PRECAUTION

To avoid contamination use clean laboratory materials.


SAMPLE

Serum/plasma (free of haemolysis)

CALIBRATION




BIBLIOGRAPHY

1. Clin, Chim, Acta 105,147-172 (1980).

2. Thefeld, W. et.al.,Dtsh.med.wschr.99,343 (1994)

SGPT 4 x 35 / 2 x 18 mL

12005021

ADL/V.01/APR 2012

Parameters Screen

Test SGPT

No

Full Name SGPT

Standard No. 2


Primary WL 340

Sec. WL

Direction Decrease

Reaction Time 4 12

Incubation Time 10

Unit U/L

Precision 0.1

R1 200 µL

R2 50 µL

Sample 10 µ L

R1 Blank 0 20000



Linearity Limit 0.2


Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low 0

High 49

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

SGPT

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Total Protein inserum or plasma.

-Direct Biuret Method

-Linearity up to 15 gm/dL.


Proteins form the major portion of dissolved substances in the plasma. They formthe basic structural components of the body. They constitute the enzymes presentin our body & also act as secondary source of energy. The other functions includedistribution of water, buffering, transport of various components, defense &coagulation of blood in our body.

Increased levels are found in dehydration & myeloma.

Decreased levels are found in liver disorders, Nephrotic syndrome, malnutrition &protein due to haemorrhage.

PRINCIPLE

Colorimetric determination of total protein based on the principle of the Biuretreaction (copper salt in an alkaline medium). Protein in plasma or serum sampleforms a blue colored complex when treated with cupric ions in alkaline solution.The intensity of the blue color is proportional to the protein concentration.

REAGENT COMPOSITION

TOTAL PROTEIN REAGENT 4 x 30 mL

Potassium Iodide 6 mmol/L

Potassium sodium tartarate 21 mmol/L

Copper Sulphate 6 mmol/L






STORAGE & STABILITY

The sealed reagents are stable up to the expiry date stated on the label, when storedat room temperature .

LINEARITY

The procedure is linear up to 15 gm/dL.

If the concentration is greater than linearity (15 gm/dL), dilute the sample withnormal saline and repeat the assay. Multiply the result with dilution factor.

NORMAL RANGE



Serum : 6.2 – 8.0 gm/dL

PRECAUTION



SAMPLE


CALIBRATION




BIBLIOGRAPHY

Gomall, A.J.Biol. Chem, 177 C (1949) 751

TOTAL PROTEIN 4 x 30 mL

12005022

ADL/V.01/APR 2012

Parameters Screen

Test TPR

No

Full Name Total Protein

Standard No. 2


Primary WL 546

Sec. WL

Direction Increase

Reaction Time 0 22

Incubation Time

Unit g/dL

Precision 0.1

R1 250 µL

R2 0

Sample 5 µL

R1 Blank 0 3000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

TOTAL PROTEIN

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of Triglycerides inserum or plasma.

-GPO-PAP methodology


-Contains LCF (Lipaemic clearing factor ) which minimizes returns


Triglycerides are simple lipids, formed in the liver by glycerol & fatty acids. Theyare transported by VLDL, LDL & constitute about 95% of fat, stored as source ofenergy in the tissue & plasma.

Increased levels are found in hyperlipidemias, diabetes, nephrotic syndrome &hypothyroidism. Increased levels are risk factor for arteriosclerotic coronary disease,peripheral vascular disease, acute pancreatitis & hyperlipoproteinaemia. Decreasedlevels are found in malnutrition & hyperthyroidism.

PRINCIPLE

Enzymatic determination of triglyceride is based on following reactions:

Lipoprotein lipase

TGL+H2O -------------------------> Glycerol + Fatty acid

Glycerol kinase

Glycerol + ATP ---------------------------> Glycerol-3-phosphate + ADP

GPO

Glycerol-3-phospahte+O2 --------------> Dihydroxyacetone phosphate

+H2O

2

POD

H2O

2+4-Aminoantipyrine+p-Chlorophenol---------------> Quinone+4H

2O

GPO = Glycereol-3-phosphate Oxidase.



AAP=aminoantipyrin

REAGENT COMPOSITION

TRIGLYCERIDES 4 x 35 mL



Potassium ferrocynate 10 mmL

Magnesium salt 17 mmol/L

4-Aminoanyipyrine 0.9 mmol/L

ATP 3.15 mmol/L

Lipoprotein lipase >1800 U/L


Glycerol -3-phosphate oxidase > 3500 U/L

Peroxidase >450 U/L





LINEARITY



NORMAL RANGE



Male 60-165 mg/dL

Female 40-140 mg/dL

PRECAUTION



SAMPLE


TRIGLYCERIDES 4 x 35 mL

12005023

CALIBRATION




BIBLIOGRAPHY

1. Schettler G. Nussel,E,Arav, Med. 10,25 (1975) J2. Jacobs,N.J. ,Van Demark,P.J. Arch Biochem. Biophy. 88,250-255 (1960)

ADL/V.01/APR 2012

Parameters Screen

Test TGL

No

Full Name Triglycerides

Standard No. 2


Primary WL 510

Sec. WL 630

Direction Increase

Reaction Time 0 20

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2 0

Sample 3 µL

R1 Blank 0 3000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

TRIGLYCERIDES

ADL/V.01/APR 2012

INTENDED USE

This reagent is intended for in vitro quantitative determination of urea in serum,plasma & urine.

-Urease / GLDH methodology



Proteins cannot be stored in human body, so excess should be broken down. Aminoacids which from the components of proteins, break down to give ammonia. Thisis toxic & so through a series of chemical reactions (urea cycle) non toxic urea isproduced & this is released into the blood which is filtered in the kidney & excretedin the urine.


Decreased levels are found in liver failure & pregnancy..

PRINCIPLE

urease

Urea + H2O --------------------> 2NH

3 + CO

2

GLDH

2 NH4+2a-ketoglutarate + 2NADH --------------> 2L-Glutamate+2NAD+ 2H

2O

REAGENT COMPOSITION

UREA U.V. R1 4 x 30 mL

Buffer (pH 7.6) 100 mmol/L

ADP 0.7 mmol/L


GLDH >1100 U/L

Urease >6500 U/L

UREA U.V. R2 2 x 16 mL

NADH 0.25 mmol/L

2-Oxoglutarate 5 mmol/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum/Plasma : 10-50 mg/dL

Urine : 20-35 gm/24 hr

PRECAUTION



SAMPLE

Serum, plasma (free of haemolysis).

(Do not use anticoagulants containing fluorides and ammonium ions)

Urine (Diluted to 1/100 with Dilled water)

Multiply the result with the dilution factor.

CALIBRATION




BIBLIOGRAPHY

1. Talke, H. and Schubert G.E.Kiln-Wocchsr 43: 174 (1965)2. Kassirer, J.P.,New Eng. J.Med. 285,385 (1971)

UREA U V 4 x 30 / 2 x 16 mL

12005024

ADL/V.01/APR 2012

Parameters Screen

Test UREA

No

Full Name Urea UV

Standard No. 2

Reaction Type Fixed time

Primary WL 340

Sec. WL

Direction Decrease

Reaction Time 3 7

Incubation Time 6

Unit mg/dL

Precision 0.1

R1 200 µL

R2 50 µL

Sample 3 µL

R1 Blank 0 10000



Linearity Limit


Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

UREA U V

ADL/V.01/APR 2012

INTENDED USE: This reagent is intended for in vitro quantitative determination ofUric acid in serum, plasma & urine.

-Uricase methodology.



Uric acid is the end product of purine metabolism. Uric acid is excreted by thekidneys.

Increased levels are found in Gout, arthritis, impaired renal functions & starvation.

Decreased levels are found in yellow atrophy of the liver.

PRINCIPLE

Enzymatic determination of Uric acid according to the following reactions.

Uricase

Uric acid + 2H2O+O

2 ----------------------> Allantoine +CO

2 +H

2O

2

Peroxidase

2H2O

2 + 4 – Aminoantipyrine +EHSPT--------------->Red quinone

EHSPT= N-Ethyl N-(2-Hydroy -3 Sulfopropyl) n-Toluidine

REAGENT COMPOSITION

URIC ACID R1 2 x 35 mL


EHSPT 1.10 mmol/L

Ferrocyanure 50 mmol/L

Amino -4-antipyrine 0.37 mmol/L


Uricase > 140 U/L





STORAGE & STABILITY


LINEARITY



NORMAL RANGE



Serum / Plasma :

Men 3.4-7.0 mg/dL

Women 2.4-5.7 mg/dL

PRECAUTION



SAMPLE

Serum, EDTA or Heparin plasma (free of haemolysis)

CALIBRATION




BIBLIOGRAPHY

1. Barham,D.,Trider P., Analyst 97,142 (1972)

2. Fossatj Clin. Chem. 26/2,227 (1980)

URIC ACID 2 x 35 mL

12602001

ADL/V.01/APR 2012

Parameters Screen

Test Uric Acid

No

Full Name Uric Acid

Standard No. 2


Primary WL 546

Sec. WL 670

Direction Increase

Reaction Time 0 38

Incubation Time

Unit mg/dL

Precision 0.1

R1 300 µL

R2

Sample 6 µL

R1 Blank 0 2000

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC Abs

Reference Screen

Low

High

Calibration Screen


Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1

Control 2

URIC ACID

ADL/V.01/APR 2012

INTENDED USEThis reagent is intended for in vitro quantitative determination of HbA1c in humanblood.Latex enhanced ImmunoturbidimetryReady to use liquid stable reagentsMultipoint calibrationDirect result (% HbA1c) from analyzerNo total Hb determination required

CLINICAL SIGNIFICANCEHbA1c is a glycated form of haemoglobin formed by the attachment of glucoseresidues in the blood to the hemoglobin molecules. In the diabetic populationwhere blood glucose levels are abnormally elevated the level of HbA1c alsoincreases. The level of HbA1c is proportional to the level of glucose in the bloodand has been widely accepted as an indicator of the mean blood glucoseconcentration in the preceeding 6-8 weeks. It is therefore a long-term indicator ofdiabetic control. For routine use HbA1c levels should be monitored every 3-4months. However in gestational diabetes and after a change in therapy it may beuseful to measure HbA1c more frequently at 2-4 week intervals.

PRINCIPLEThe principle of the test is latex agglutination method that measures the ratio ofhemoglobin A1c that occupy in a total hemoglobin in the whole blood. The sample(hemolysis sample ) is added to the unsensitized latex particles, and the surfaces ofthe latex adsorbs a total hemoglobin in the sample. Anti human HbA1c mousemonoclonal antibody complex agglutinates by anti-mouse IgG goat antibody. At thistime, the amount of agglutination caused depends on the amount of HbA1c thatadsorbs the surface of the latex, this this agglutination is measured as a turbidity. theconcentration of HbA1c (%) in the sample is determined by referring to the calibrationcurve obtained by the same test of diluted standard solutions.

REAGENT COMPOSITIONHbA1c R1 1 x 15 mL/2 x 15 mLLatexHbA1c R2 1 x 5 mL / 2x 5 mLAnti-human HbA1c mouse monoclonal antibodyAnti-mouse IgG goat antibodyHbA1c R3 1 x 32 mL / 1 x 63 mLHaemolysis ReagentHbA1c DIRECT CALIBRATOR 4 x 0.5 mLHbA1c 4 level calibrator (lyophilized).

STORAGE AND STABILITYThe sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C, protected from light. Once opened the reagent is stable upto four weeks, ifcontamination is avoided. Recalibration is recommended after 30 days. DO NOT FREEZECalibrator : Reconstitute the calibrator with 0.5mL distilled water. It is stable for 30days at 2-8oC . DO NOT FREEZE.

LINEARITY RANGEThe reagent is linear upto 13% (NGSP)

REFERENCE RANGEIt is recommended that each laboratory establish its own reference values.The following value may be used as guide line.Reference normal value (NGSP): 4.6% -6.2%

PRECAUTIONTo avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent and standard bottles immediately after use.Avoid direct exposure of working reagent to light.

SAMPLEWhole blood, collected with EDTATo determine HbA1c a heamolysate must be prepared for each sample1. Dispense 1mL hemolysis reagent into a tube.2. Add 20 μLof well-mixed whole blood and mix.3. Allow to stand for 5 minutes or until complete lysis is evident.

Follow the same procedure with controls and calibrator.

GENERAL SYSTEM PARAMETER FullyAutoMode of Reaction End pointSlope of reaction IncreasingWavelength 660 nmTemperature 370CCalibrator Concentration As on vial labelLinearity 13%Blank Reagent blankSample volume 7.5 μLReagent 1 volume 180 μLReagent 2 volume 60 μLCuvette 1 cm light pathLABORATORY PROCEDURE FOR FULLY AUTO

Blank Calibrator Sample/controlHbA1c R 1 180 μL 180 μL 180 μLCalibrator - 7.5μL -Hemolysate(sample/control) - - 7.5μLMix & incubate for 5 min at 370C.HbA1c R 2 60 μL 60 μL 60 μLMix and incubate for 5 min at 37oC and read absorbance(A) at 660 nm.

CALCULATIONCalibration curveCalculate the Abs of calibrators = Abs calibrator – Abs Blank. Plot the D Abs of eachcalibrator versus assigned concentration (HbA1c %) on a linear graph paper. HbA1cresults according to NGSP for the samples and controls are determined using theprepared calibration curve.Calculate

Abs of sample ie abs Sample - abs Blank .HbA1c % in the sample is calculated by interpolation of Abs of sample on thecalibration curve. For calculation of results according to IFCC, use IFCC calibratorvalues (see calibrator insert), or use following equation.NGSP = (0.915 x IFCC) + 2.15

INTERFERENCESNo interference upto :Ascorbic acid 50 mg/dLBilirubin 40mg/dLIntra lipid 3000 mg/dL


BIBLIOGRAPHY1. Engbeak, F., et al. Clin chem.35 p. 93-97 (1989)2. American Diabetes Association : Clinical practice recommendations (position

statement). Diabetes care 24 (suppl.1) S33-S55, (2001).3. Tietz, N.W. Textbook of Clinical Chemistry, W.B. Saunders Company, p.794 -795

(1999).

HbA1c DIRECT WITH CALIBRATOR 1 x 15 /1 x 5/ 1x 32 /4 x0.5mL11835001

ADL/V.02/July 2013

2 x15/2 x5 /1x 63/4x0.5mL11835002

xxxxxxxxxxxxxxxxxxxxxx+91 484 3120 002

INTENDED USE: This reagent is intended for in vitro quantitative determination ofHbA1c in human blood.-Latex enhanced Immunoturbidimetry-Multipoint calibration-Direct result in % HbA1c from analyzer-No total Hb determination requiredCLINICAL SIGNIFICANCEHbA1c is a glycated form of haemoglobin formed by the attachment of glucoseresidues in the blood to the hemoglobin molecules. In the diabetic populationwhere blood glucose levels are abnormally elevated the level of HbA1c alsoincreases. The level of HbA1c is proportional to the level of glucose in the bloodand has been widely accepted as an indicator of the mean blood glucoseconcentration in the preceeding 6-8 weeks. It is therefore a long-term indicator ofdiabetic control. For routine use HbA1c levels should be monitored every 3-4months. However in gestational diabetes and after a change in therapy it may beuseful to measure HbA1c more frequently at 2-4 week intervals.

PRINCIPLEThe principle of the test is latex agglutination method that measures the ratio ofhemoglobin A1c that occupy in a total hemoglobin in the whole blood. The sample(hemolysis sample ) is added to the unsensitized latex particles, and the surfaces ofthe latex adsorbs a total hemoglobin in the sample. Anti human HbA1c mousemonoclonal antibody complex agglutinates by anti-mouse IgG goat antibody. At thistime, the amount of agglutination caused depends on the amount of HbA1c thatadsorbs the surface of the latex, this this agglutination is measured as a turbidity. theconcentration of HbA1c (%) in the sample is determined by referring to the calibrationcurve obtained by the same test of diluted standard solutions.REAGENT COMPOSITIONHbA1c R1 1 x 30 mLLatexHbA1c R2 1 x 10 mLAnti human HbA1c mouse monoclonal antibodyAnti-mouse IgG goat antibodyHbA1c R3 2 x 63 mLHaemolysis ReagentReagent required but not providedAgappe HbA1C Direct Multicalibrator ( 11604001)STORAGE AND STABILITYThe sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C, protected from light. Stability in the instrument is four weeks, ifcontamination is avoided. Recalibration is recommended after 30 days. DO NOT FREEZELINEARITYThe reagent is linear up to 13% (NGSP)REFERENCE RANGEIt is recommended that each laboratory establish its own reference values.The following value may be used as guide line.Reference normal value (NGSP): 4.6% -6.2%PREPARATION AND STABILITY OF WORKING REAGENTReagent 1 & Reagent 2 are ready to use.

CALIBRATIONAgappe HbA1C Direct Multicalibrator ( 11604001) is recommended for calibrationof this assay. Reconstitute the calibrator with 0.5 mL distilled water and it will be stableup to 30 days at 2-80C. Do not freeze.Quality ControlIt is recommended to use Agappe HbA1C Control Level 1&2 (11625002) to verifythe performance of the assay. Each laboratory has to establish its own internal qualitycontrol scheme and procedure for corrective action, if controls do not recover within the acceptable tolerance.PRECAUTIONTo avoid contamination, use clean laboratory wares. Use clean, dry disposable pipettetips for dispensing. Close reagent and standard bottles immediately after use.Avoid direct exposure of working reagent to light.

SAMPLEWhole blood, collected with EDTATo determine HbA1c a heamolysate must be prepared for each sample1. Dispense 1mL hemolysis reagent into a tube.2. Add 20 μL of well-mixed whole blood and mix.3. Allow to stand for 5 minutes or until complete lysis is evident.


INTERFERENCESNo interference up to :Ascorbic acid 50 mg/dLBilirubin 40 mg/dLIntralipid 3000 mg/dLIt has been reported that results may be inconsistent in patients who have thefollowing conditions: opiate addiction, lead poisoning, alcoholism, and ingestion oflarge doses of aspirin. Elevated HbF levels may lead to under estimation of HbA1c.BIBLIOGRAPHY1. Engbeak, F., et al. Clin chem.35 p. 93-97 (1989)2. American Diabetes Association : Clinical practice recommendations (position

statement). Diabetes care 24 (suppl.1) S33-S55, (2001).3. Tietz, N.W. Textbook of Clinical Chemistry, W.B. Saunders Company, p.794 -795

(1999).

HbA1c DIRECT 1 x 30/1 x 10/2 x 63 mL12005057

ADL/V.02/28.06.2013

xxxxxxxxxxxxxxxxxxxxxx+91 484 3120 002

Parameters Screen

Test HbA1c D

No

Full Name HbA1c Direct

Standard No. 5


Primary WL 670

Sec. WL

Direction Increase

Reaction Time -1 19

Incubation Time 12

Unit %

Precision 0.01

R1 180 μL

R2 60 μL

Sample 7.5μL

R1 Blank

Mixed Rgt.Blank


Linearity Limit

Substrate Limit

Factor

Prozone chek

q1 q2 q3 q4

PC

Refence Screen

Low *

High *


Calibration Screen

Rule Spline

Sensitivity

Replicates 1

Interval ( day) 0

Difference Limit

SD

Blank Response

Error Limit


QC Screen

Control 1 *

Control 2 *

HbA1c DIRECT

ADL/V.01/28.06.2013

AGAPPE DIAGNOSTICS LTD.

Agappe Hills, Dist. Ernakulam, Kerala, India - 683 562Customer Care No. [Toll Free] : 1800 425 9800 | [email protected] | www.agappe.com

ISO 9001 : 2008ISO 13485 : 2003

IVD LOT LOT NUMBERSYMBOLS USED ON THE LABELS : SEE PACKAGE INSERT FOR PROCEDUREIN VITRO DIAGNOSTIC USE MANUFACTURER S ADDRESS’ TEMPERATURE LIMITEXPIRY DATEMANUFACTURING DATE

VERSION No. :

INTENDED USE: This reagent is intended for in vitro quantitative determination ofHbA1c in human blood.Latex enhanced ImmunoturbidimetryMultipoint calibrationDirect result in % HbA1c from analyzerNo total Hb determination requiredCLINICAL SIGNIFICANCEHbA1c is a glycated form of haemoglobin formed by the attachment of glucoseresidues in the blood to the hemoglobin molecules. In the diabetic populationwhere blood glucose levels are abnormally elevated the level of HbA1c alsoincreases. The level of HbA1c is proportional to the level of glucose in the bloodand has been widely accepted as an indicator of the mean blood glucoseconcentration in the preceeding 6-8 weeks. It is therefore a long-term indicator ofdiabetic control. For routine use HbA1c levels should be monitored every 3-4months. However in gestational diabetes and after a change in therapy it may beuseful to measure HbA1c more frequently at 2-4 week intervals.PRINCIPLEThe kit is based on latex agglutination method that measures the ratio of hemoglobinA1c that occupy in a total haemoglobin in the whole blood. The sample is added tothe unsensitization latex particles, and the surfaces of the latex adsorbs a totalhaemoglobin in the sample. Anti -human HbA1c mouse monoclonal antibody is toreact and anti-human Hba1c mouse monoclonal antibody complex agglutinates byanti-mouse IgG goat anti body. At this time the amount of agglutination causeddepends on the amount of HbA1c that adsorbs the surface of the latex, thisagglutination is measured as a turbidity. The concentration of HbA1c(%)in the wholeblood is calculated from a standard curve.REAGENT COMPOSITIONHbA1c R1 1 x 26 mLLatexHbA1c R2 1 x 9 mLAnti-human HbA1c mouse monoclonal antibodyAnti mouse IgG goat antibodyHbA1c R3 1 x 63 mLHaemolysis ReagentReagent required but not providedAgappe HbA1C Direct Multicalibrator ( 11604001) STORAGE AND STABILITYThe sealed reagents are stable up to the expiry date stated on the label, when storedat 2- 80C, protected from light. Stability in the instrument is four weeks, ifcontamination is avoided. Recalibration is recommended after 30 days. DO NOT FREEZEPRECAUTIONClose reagent and standard bottles immediately after use. Keep the reagent back tospecified temperature after each useAvoid direct exposure of reagent to light.Avoid freezing of reagents and refrigerate them at proper temperature. Shake R1and R2 reagent well before use.LINEARITYThe reagent is linear up to 13% (NGSP)REFERENCE RANGEIt is recommended that each laboratory establish its own reference values.The following value may be used as guide line.Reference normal value (NGSP) : 4.6 - 6.2 %PREPARATION AND STABILITY OF REAGENTReagent 1, reagent R2 and reagent R3 are ready to use.

CALIBRATIONAgappe HbA1C Direct Multicalibrator (11604001) is recommended for calibration ofthis assay. Reconstitute the calibrator with 0.5 mL distilled water and it will be stableup to 30 days at 2-80C. Do not freeze.Quality ControlIt is recommended to use Agappe HbA1C Control level 1&2 (11625002) to verify theperformance of the assay. Each laboratory has to establish its own internal qualitycontrol scheme and procedure for corrective action, if controls do not recover within the acceptable tolerance.SAMPLEWhole blood, collected with EDTATo determine HbA1c a heamolysate must be prepared for each sample1. Dispense 1mL hemolysis reagent into a tube.2. Add 20 μL of well-mixed whole blood and mix.3. Allow to stand for 5 minutes or until complete lysis is evident.


INTERFERENCESNo interference upto :Ascorbic acid 50 mg/dLBilirubin 40 mg/dLIntralipid 3000 mg/dL

It has been reported that results may be inconsistent in patients who have thefollowing conditions: opiate addiction, lead poisoning, alcoholism, and ingestion oflarge doses of aspirin. Elevated HbF levels may lead to under estimation of HbA1c.BIBLIOGRAPHY1. Nathan, D.M., Clin, Chem. 29, pp.466-469 (1983)2. Engbeak, F., et al. Clin chem.35 pp. 93-97 (1989)3. American Diabetes Association : Clinical practice recommendations (position

statement). Diabetes care 24 (suppl.1) S33-S55, (2001).4. Tietz, N.W. Textbook of Clinical Chemistry, W.B. Saunders Company,

p.794 -795 (1999).

HbA1c DIRECT 1 x 26 mL/1 x 9 mL/1x 63 mL

12013042

ADL/V.01/July 2013

Page 1Test (Name) HbA1cDecimal 2Unit %Reac. Method EndBlank R BlankWave length I 670Wave length II -Test Time 270 - 300Sample 8 μLR1 210 μLR2 70 μLCalculation Method Spline

Mispa Mini Assay Parameter

Procedure_Catalogue-Phoenix-All_Cobined.pdf - Bio Group

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