Antibody Titration Test Method - NSW Health Pathology

Antibody Titration Test Method NSWHP_PR_063

of 17 Approver: Director Clinical Operations, Version Number: V1.0, Publication Date: 27/10/2021 This document is subject to

change and a printed copy may not be up to date. The current version is only available online in the NSW Health Pathology Policy Library

www.pathology.health.nsw.gov.au

1. Purpose

This document outlines the Bio-Rad CAT methodology for performing antibody titration testing across

NSW Health Pathology Laboratories.

1.1. Principle of Assay

Titration studies are semi-quantitative and undertaken to support the prediction and management of

Haemolytic Disease of the Fetus and Newborn (HDFN), ABO mismatched organ transplants and

Allogeneic Transplant of ABO mismatch Bone Marrow and Peripheral Blood Stem Cell Transplants. A

titre is a measure of the strength of an antibody. Titration values can help quantify the relative antibody

concentration in the plasma, or the relative strength of antigen expression on red cells. In general, an

antibody reacting at a high titre (e.g. 128) will cause greater red cell destruction than an antibody

reacting at a low titre (e.g. 2).

The titre is obtained by testing serial dilutions of the antibody against appropriate red cells and a titre

value assigned based on the highest dilution of the antibody still causing a positive reaction of a

particular strength, depending on the antibody being titred. The haemagglutination titre is expressed as

the reciprocal of the highest serum dilution in which agglutination is observed as a score of 1. At Royal

Prince Alfred Hospital the renal titre end point is 2.

Titres have wide application in the Transfusion laboratory:

• Monitoring the antibody level in an antenatal patient

• Monitoring antibody mediated rejection (ABMR) following ABO mismatched organ transplants.

• Anti-A and anti-B titres are performed pre and post-pheresis prior to the infusion of mismatched bone

marrow or stem cells for transplant.

• Detection of underlying alloantibodies in patients with autoantibodies

• Identification and investigation of HTLA antibodies.

Antenatal Titres

Antibody titres are performed on antenatal patients where potentially clinically significant IAT reactive antibodies are detected to estimate the risk of HDFN. Anti-D, c and K are most commonly associated with HDFN but anti-C, Cw, E, e, k, Fy, Jk, M, S are potentially significant and require titration. An increase in the antibody titre may relate to an increase in the concentration of antibody which could reflect an increase in risk to the fetus. However previous history is not indicative of the severity of disease for anti-K antibodies.

After identification and titration, plasma is separated and stored frozen for parallel titration with

subsequent samples. This will ensure that any apparent change in titre is not due to technique or

reagent variation. Anti-c and immune anti-D are referred to Australian Red Cross Lifeblood (ARCL)

reference laboratory for antibody quantitation.

ABO Titres

The ABO antibody that is titrated in the patient has the specificity of the ABO antigen being transplanted

from the donor, e.g. If the patient is Group O and the Donor is Group A, then Anti-A in the patient will be

titred. ABO Tires are performed using Bio-Rad ID-Diacell ABO (A1, A2, B Cells) Pooled Red Cells for

Reverse Serum Grouping as follows:

https://intranet.pathology.health.nsw.gov.au/toolbox/policies-and-procedures/policies





1. Renal transplant patients having an ABO-incompatible transplant.

Pretransplant workup titre against potential donor, A1, A2 or B cells depending on the blood group or

group A sub-type of the donor.

Peri and Post-transplant titre against A1, A2 or B cells depending on the blood group or the group A

sub-type of the donor.

2. Anti-A titres are performed on all group O and group B pre-liver transplant patients.

3. Haematology Patients undergoing Allogeneic Transplant of ABO Major mismatch Haemopoietic

progenitor cells (HPC). Titre against A1 or B cells as indicated.

Patients undergoing an ABO incompatible renal transplant often require titres to be performed. Titres are

performed pre transplant to monitor the response to treatment and post transplant to aid in the

assessment of the success of the transplant. The titres are performed for Anti-A1, Anti-A2 and Anti-B,

based upon the blood group of the patient and potential donor.

Prior to Allogeneic Transplant of ABO major mismatch HPC donor cells plasmapheresis may be

performed if the ABO titre of the patient is high to reduce or remove the ABO IgM antibodies. This will

minimise the destruction of ABO Major mismatch transplanted cells which may be destroyed if high titres

of ABO antibodies are present. A post plasmapheresis sample, prior to the transplant date may be

received to measure the success of plasmapheresis in the removal of these IgM antibodies. Further

plasma exchanges may occur if the desired ABO titre has not been achieved. Donor ABO Titres (anti-A

and/or anti-B titres) are performed for minor mismatch. If the donor titre is high plasma reduction will be

performed on bone marrow or peripheral blood stem cell collection.

1.2. Scope

This test method applies to all NSW Health Pathology scientific and technical staff involved in

pretransfusion laboratory practice.

1.3. Test Frequency and Expected Turnaround Time

Performed on request during laboratory hours.

Refer to local laboratory procedures for expected turnaround times.

1.4. Definitions

ABOi: Blood group incompatibility

IgM: Immunoglobulin M

IgG: Immunoglobulin G

HDFN: Haemolytic Disease of the Fetus and Newborn

ABMR: Antibody Mediated Rejection

IAT: Indirect Antiglobulin Test

CAT: Column Agglutination Technology

EDTA: Ethylenediaminetetraacetic acid

LIS: Laboratory Information System






RCPA QAP: Royal College of Pathologists of Australasia Quality Assurance Program

ARCL: Australian Red Cross Lifeblood

2. Method Specifications

2.1. Potential Sources of Assay Variability

• Bacterial or other contaminations of reagents can cause false positive or false negative results, look

for turbidity. Repeat test with new reagents.

• Fibrin in the plasma or red cell aggregates in the red cell suspension may trap non-agglutinated cells

at the top of the gel while most of the cells are on the bottom of the microtube after centrifugation.

Remove fibrin and repeat test.

• Elevated white cell counts may cause false positive reactions. Consult with laboratory senior.

• Too heavy or too light red cell suspensions may cause discrepant results.

• Correct pipetting technique must be used to avoid contact of the pipette tip with the contents of the

microtubes to prevent carryover.

3. Pre-Analytical

3.1. Primary Specimen Type e.g. blood, urine, tissue

Venous blood collected aseptically.

3.2. Specimen containers including any additives, anticoagulants, buffers, acid

• An anticoagulated EDTA (pink or purple top) specimen tube centrifuged as per local policy prior to

testing.

• Other tube types that may be accepted for testing include citrate (blue top) clotted blood (red top) and

CPD-A centrifuged as per local policy prior to testing.

• Lithium heparin (green top) and serum gel tubes are not acceptable due to the potential for the gel to

absorb antibodies resulting in false negative results.

Note: Throughout the document reference to plasma and serum is interchangeable.

3.3. Assay Validation and Verification

Validation and Verification documentation of the test method is reviewed and signed by the Supervising

Pathologist and retained in the Quality Management System. For further information refer to Verification

Procedure for the Bio-Rad IH 1000, IH 500 and Bio-Rad Saxo Analysers NSWHP_PR_046.

3.4. Sensitivity, Specificity of detection

An anti-D titre using Securacell 1 must be performed daily when a titre is performed. The expected

titration endpoint of Securacell 1 is 32 (+/-1). If this result is not achieved all testing including the

Securacell 1 quality control is repeated.

3.5. Patient preparation e.g. fasting, time after last dose – if applicable






Not Applicable

3.6. Specimen Rejection Criteria

A specimen will be rejected for testing if:

• The specimen does not meet the criteria outlined in Labelling Requirements for Pathology and Forensic

Specimens NSWHP_PD_023. It is not necessary that the additional requirements for Transfusion

specimens are met.

• The specimen is insufficiently filled. Refer to local recollection procedures.

• The specimen is grossly haemolysed. Refer to local recollection procedures.

3.7. Specimen Transport Requirements – frozen, room temp, on ice, other

Specimens may be transported at room temperature or 2-8oC.

3.8. Specimen Stability, Storage and Retention

Specimens should be tested within 24 hours of collection wherever possible, and testing completed

within 48 hours when kept at room temperature (18-25ºC) and up to 7 days when stored between 2-8oC.

Specimens stored at 2-8oC are not used for serological testing beyond 7 days from the date of collection.

If specimens are required to be routinely tested after 7 days, they must be separated, and the plasma

stored frozen at -20 ºC for up to 3 months. If parallel testing with a sample older than 3 months is

required, an anti-D titre with Securacell 1 can be performed in parallel as a control. Refer to local

laboratory procedures for specimen retention time.

4. Reagents, Supplies and Equipment

4.1. Required Reagents and Reagent Source

DiaClon ID-Card LISS/Coombs: Indirect and Direct Coombs Test (Product ID 50531)

ID-Diacell (I-II-III) Test cell reagents for antibody screening (Product ID 45184)

ID-DiaPanel (Set of 11 vials) Test cell reagents for antibody identification- IAT (ID 45161)

ID-Diacell ABO (A1, A2, B Cells) Pooled Red Cells for Reverse Serum Grouping (ID 45082)

ID-Diluent 2 modified LISS for red cell suspensions (Product ID 05761)

ID-Titration racks- containing 60 dilution wells for use on the IH-500 (Product ID 05770)

ID-Titration Solution- Bovine Serum Albumin (BSA) based diluent (Product ID 05780)

0.9% Buffered Saline Solution pH 7.1

Securacell

4.2. Reagent Preparation

Not Applicable






4.3. Reagent Stability and Storage

• ID-Diacell (I-II-III), ID-DiaPanel and ID-Diacell ABO red cell reagents store at 2-8ºC and do not store

near any heat, air conditioning sources or ventilation outlets. Red cell reagents must be gently

resuspended and at room temperature (18-25 °C) when used for manual testing. Reagents are used

until the expiry date as indicated by the manufacturer. Refrigerated reagents stored onboard the IH-

500 analyser are stable for 7 days.

• ID-Titration Solution store at 2-8ºC and do not store near any heat, air conditioning sources or

ventilation outlets. The Titration Solution is used until the expiry date as indicated by the

manufacturer, once opened the expiry of the vial is 4 weeks.

• DiaClon ID-Cards store at 18-25ºC. Inspect cards before use ensuring that buffer covers the gel and

that there are no bubbles in the gel. DiaClon ID-Cards are used until the expiry date as indicated by

the manufacturer. ID-Cards stored onboard the IH-500 analyser are valid for 21 days.

• Securacell store at 2-8ºC and do not store near any heat, air conditioning sources or ventilation

outlets. Reagents are used until the expiry date as indicated by the manufacturer.

4.4. Laboratory Equipment

Automated Bio-Rad CAT method

Testing is performed on the Bio-Rad IH-500 fully automated system for immunohaematology

diagnostics.

Manual Bio-Rad CAT Method

The Bio-Rad Saxo ID-Reader II is a semi-automated system for immunohaematology diagnostics and

relies heavily on the technician correctly following testing procedures. Manually pipetted gel cards are

incubated, centrifuged, the reactions read by the integrated camera and the results interpreted by the

interpretation software.

• ID-Dispenser (for the ID-Diluent 2)

• Pipette and Pipette tips

• ID-Working table

• Test tubes

• ID-Incubator L

• ID-Centrifuge L (back up centrifuge)

4.5. Quality Control and QC Acceptance Criteria

Pre acceptance testing is performed on all new batches of ID-Diacell (I-II-III), ID-DiaPanel, ID-Diacell

ABO red cell reagents and ID-Cards before being placed into general use. Refer to Pre Acceptance

Testing of Red Cell Reagents NSWHP_PR_051 and Pre Acceptance Testing Transport and Storage of

Bio-Rad ID-Cards and Solutions NSWHP_PR_050 for further information.

Quality control for the Forward/Reverse Cards, Group Check Cards, LISS/Coombs ID-Cards, and ID-

Diacell (I-II-III) are performed daily using QC material. An anti-D titre using Securacell 1 is performed

daily for each technician when performing titres manually and daily for the analyser if testing is

automated. The expected titration endpoint of Securacell 1 is 1:32 (+/-1). QC results will be accepted for


https://intranet.pathology.health.nsw.gov.au/toolbox/policies-and-procedures/policies?dfaction=search&df1630_6=NSWHP_PR_051






validation if the expected titration endpoint is reached. If this result is not achieved, all testing (including

any patients run in parallel with the QC anti-D titre) must be repeated, ensuring the appropriate R2R2

screening cell is selected and that Securacell 1 has not expired. Any discrepancies must be investigated,

report unresolved QC out of range to laboratory senior. Patient antibody titre results will not be accepted

until QC results are acceptable.

4.6. Calibration Procedures

The calibration and preventative maintenance of the Bio-Rad Saxo ID-Reader II, ID-Incubator L and ID-

Centrifuge L is performed annually by qualified Bio-Rad engineers.

The calibration of the IH-500 is performed at 6 monthly preventative maintenance services by qualified

Bio-Rad engineers.

Pipette checks are performed 6 months.

Centrifuge calibrations are performed annually.

4.7. Quality Assurance Programs

NSWHP Laboratories are enrolled in RCPA QAP programmes, laboratories performing antibody titres

are enrolled in the Antibody Titre module. NSWHP Transfusion Laboratories participate in an

interlaboratory sample exchange program for tests in scope not covered by RCPA QAP. Refer to local

Quality Assurance standard operating procedures for further information. All staff must participate in the

quality assurance program and undertake at least two exercises per year to ensure their gradings are

correct and that all positive tests are being detected.

5. Method Detail

5.1. Full Procedure

Automation on the Bio-Rad IH-500 is the preferred method. Results for patient testing performed on

the Bio-Rad Saxo ID-Reader and IH-500 are stored in the IH-Com database along with the patient

details, batch numbers and expiry dates of reagents, cells, ID-Cards and Diluents. All staff are allocated

a unique user ID and password which allows traceability of the user actions performed at all stages of

testing.

Standard Laboratory Procedure

1. Perform routine antibody identification or for ABO titration requests check the history of the patient in the LIS to determine which ABO antibody requires titration. For all titres check the previous titre result (if available).

2. For antenatal testing locate the frozen aliquot of a previous sample (if available), allow to thaw prior to testing in parallel with current sample.

3. Order Antibody Titration in IH-Com for tests not ordered in the LIS.

4. All results are to be printed from IH-Com (Print, Results and Protocols), ensuring the Securacell 1 has been performed and the result is printed with all patient samples.

Refer to:

• Bio-Rad Saxo ID-Reader II NSWHP_PR_057, for instructions on how to assign, centrifuge and read

ID-Cards







• Bio-Rad IH-500 Immunohaematology Analyser NSWHP_PR_055 for information on processing

specimens on the analyser.

• Bio-Rad IH-Com Data Management Software NSWHP_PR_054 for instructions on how to order tests

and review results in IH-Com.

5.2. Ordering Anti-D Quality Control – Securacell 1

An anti-D titration of a known endpoint must be performed each day a titre is performed using Securacell

1 on the IH-500. If the antibody titration is being performed manually with the Saxo ID-Reader II

Securacell 1 must be performed manually each day by each technician performing an antibody titration.

In IH-Com, select Work list tab, select ‘Quick Request’.

From the drop down list select the test ‘Anti-D titre’

Scan the barcode on Securacell 1.

Depending on the method of testing, either follow the steps for 5.7 Antibody Titre by manual CAT

method or 5.8 Antibody Titre by automated CAT method.

5.3. Ordering an Antibody Titration in IH-Com

At the Main (Analyser) screen, select ‘Data Management’.

Select the ‘Work List’ tab.

Right Click and select ‘Quick Request’.

Select the test from the dropdown list.

The syntax of the titration test is:

Titration [gel card name], [dilution level], [incubation temperature] x [reagent code]

e.g. Titration LISS/Coombs (5053), 1-6, 37ºC x 0609

The reagent code is the individual cell number associated with each reagent bottle and is the first

4 digits on the barcode of the reagent bottle. This unique reagent code is displayed in IH-Com

when the respective reagent red cells are onboard the IH-500. It is important to take care when

selecting the test as the reagent code identifies the red cells to be titred.

A titration can be ordered on any of the following reagent red cells for a 1:32 or 1:2048 dilution.

a. Individual cells from 0.8% ID-DiaCell I-II-III

b. Individual cells from 0.8% ID-DiaPanel

c. A1, A2 or B cells from 0.8% ID-Diacell ABO

Refer to local laboratory procedure at Royal Prince Alfred Hospital for ABO incompatible renal

transplant workup and procedure to titre donor red cells against the recipients plasma

Scan the sample barcode, IH-Com may identify the patient, A window box will open with ‘The

sample number is already in use for another patient: Surname,Firstname,Dob (Sex)[MRN}’

select ‘Yes’ if this is the correct patient.

Click ‘Order Now’ and ‘Close’.








5.4. Ordering an Antibody Titration in IH-Com while reviewing the antibody results:

This procedure cannot be used for ABO titrations

In IH-Com, under the Results tab, review the correct patient’s antibody screen and panel results

in the Result verification screen.

If a titration is possible on the header of the positive well, a small tick box will be present, select

this box for the antibody titration to be performed.

Confirm the request by selecting the ‘Order Titration’ button at the bottom of the

screen.

The selection is displayed with a message for acknowledgement. Confirm the message by

selecting ‘OK’.

5.5. Selection of Test Cells for Antenatal Patients

A constant source of cells, that is a fixed antigen site density, eliminates one major variable which can

influence the results of titration. For anti-D titres R2R2 cells are generally used as they have the strongest

expression of the D antigen the cell of choice is cell II from 0.8% ID-DiaCell I-II-III.






If the patient has additional alloantibodies, select R2R2 cells for the Anti-D titre, ensuring it is negative for

the additional alloantibodies present. Ensure selected homozygous cells are reading consistently with

other homozygous cells in the panel.

Rh antibodies

Single Antibody Cells Required Multiple Antibodies Cells Required

Anti-D R2R2 Anti-D and anti-C R1R1 or R2R2

Anti-C R1R1 Anti-D and anti-E R2R2

Anti-E R2R2

Anti-e R1R1

Anti-c R2R2

Other Atypical Antibodies

For other antibodies, use a single homozygous positive cell for the titre being performed

from the 0.8% ID-Panel. Anti-K is an exception where homozygous expression is rare

and not found routinely in panel cells. If the titre has been previously performed using

cells heterozygote for the antigen being titred then it is important to continue with

heterozygous cells.

e.g. for Anti-Fya Fya+b- (homozygous cell)

Antibody Mixtures

For mixtures of antibodies, cells selected to estimate the strength of each antibody may

be used by selecting cells homozygous for one of the antibodies and negative for the

other.

e.g. Where anti-K and anti-Fya is present Titrate against K-, Fya+b- cells and K+, Fya-

cells

5.6. Selection of Test Cells for ABO Titrations

The selection of cells will be depending upon the antibody being titred, using A1, A2 or B cells from 0.8%

ID-Diacell ABO. If the patient has an alloantibody, the cells used for the titre must also be antigen

negative. The antibody being titred is determined by the antibody the recipient has but the donor lacks

which may affect recipient donor compatibility.






Titres can be ordered by requesting a 1:2048 titration

Test Code including Individual Cell Number Cell

Titration LISS/Coombs (5053) 1-12, 37oC, TLIS3_0608 I

ID-DiaCell I-II-III 0.8% Titration LISS/Coombs (5053) 1-12, 37oC, TLIS3_0609 II

Titration LISS/Coombs (5053) 1-12, 37oC, TLIS3_0610 III

Titration LISS/Coombs (5053) 1-12, 37oC, TLIS3_0617 1

ID-Panel 0.8%
















Titres can be ordered by requesting a 1:32 titration

Please note, the titre endpoint must be followed by two negative wells.


Titration LISS/Coombs (5053) 1-6, 37oC TLIS1_0608 I

ID-DiaCell I-II-III

0.8% Titration LISS/Coombs (5053) 1-6, 37oC TLIS1_0609 II

Titration LISS/Coombs (5053) 1-6, 37oC TLIS1_0610 III

Titration LISS/Coombs (5053) 1-6, 37oC TLIS1_0617 1

ID-Panel 0.8%
















For patients requiring an ABO titration, select one of the following options.


Titration LISS/Coombs (5053) 1-6, 37oC, TLIS1_0601 A1




Titration LISS/Coombs (5053) 1-6, 37oC, TLIS1_0603 B

Titration LISS/Coombs (5053) 1-12, 37oC, TLIS3_0603 B

Titration Coombs Anti-IgG (5053) 1-6, 37oC x 0601 A1




Titration Coombs Anti-IgG (5053) 1-6, 37oC x 0603 B

Titration Coombs Anti-IgG (5053) 1-12, 37oC x 0603 B

5.7. Antibody Titre by manual CAT method

The manual CAT method is performed using the Saxo ID-Reader II, refer to Bio-Rad Saxo ID-Reader II

NSWHP_PR_057 for instructions on how to assign ID-Cards and print a barcode label.

Prepare a twofold serial dilution:

Label a row of test tubes 2 to 12.

Pipette 200 µL of ID-Titration Solution to tubes 2 to 12.

Add 200 µL of patient plasma to tube two and mix thoroughly by aspirating and dispensing the

contents with a fresh pipette tip.

Pipette 200 µl of the dilution from tube two (1:2 dilution) to the tube three (1:4 dilution), change

pipette tips and mix tube three thoroughly.

Continue serial dilution of the patient’s plasma by repeating this procedure through to tube twelve

ensuring each tube is mixed thoroughly with a new pipette tip before transfer to the next tube to

avoid carry over of plasma.

• The titre can be extended if it is expected to exceed 1/2048 (12 tubes).







• The sample volume can be adjusted according to the availability of sample, if a different

sample volume is used adjust the volume of the diluent accordingly.

Tube 1 2 3 4 5 6 7 8 9 10 11 12

Dil’n 1:1 1:2 1:4 1:8 1:16 1:32 1:64 1:128 1:256 1:512 1:1024 1:2048

Perform Antibody Titration

Label a DiaClon LISS/Coombs ID-Card(s) with a barcode with patient and reagent details.

Remove the aluminium foil from as many microtubes as required holding the ID-Card in the upright

position.

Pipette 50 µL of the 0.8% reagent red cells to each mictotube of the LISS/Coombs cards. Ensuring

the cell is a homozygous positive cell for antibody being titred. Refer to 5.5 and 5.6 for Selection of

Test Cells.

Starting from the highest dilution, pipette 25 µL of the contents of each master dilution tube to the

corresponding microtube in the LISS/Coombs card. For tube 1 (1: dilution) add 25 µL of (neat)

patient plasma.

Incubate the ID-Card(s) for 15 minutes at 37ºC in the ID-Incubator L.

Centrifuge and read the ID-Card(s) using the Saxo-ID-Reader II.

Refer to 6.1 Result Interpretation.

Refer to LIS standard operating procedures for result entry details.






5.8. Antibody Titre by automated CAT method

The test is performed on the Bio-Rad IH-500 using the same volumes and same dilutions as outlined in

5.7 Antibody Titre by manual CAT method. Refer to 5.3 and 5.4 Ordering an Antibody titre in IH-Com for

instructions on how to request a test not ordered in the LIS. Once the testing has been requested in IH-

Com place the patient sample in the sample rack and load onto the Bio-Rad IH-500 analyser. Ensure ID-

Titration solution (at least 2 mL), ID-Titration racks and required test red cells have been loaded on

board the IH-500 via the left drawer, each titre will use 6 or 12 microtubes depending on the test being

requested. All blood groups and antibody testing will have priority over titration requests.

5.9. Result Out-Of-Range / Reflex Testing / Supplementary Testing

Not Applicable

5.10. Calculations

Not Applicable

6. Reporting Result

Refer to LIS standard operating procedures for result entry details.

6.1. Result Interpretation and choice of Coded Comments

Positive: Agglutinated cells forming a red line on the surface of the gel or dispersed in the gel.

Negative: Compact button of cells on the bottom of the microtube.

Reaction Grading for Bio-Rad CAT Testing

The titre is the reciprocal of the highest plasma dilution, which gives a strength of 1+ reaction (the

exception is the renal titre end point at Royal Prince Alfred Hospital which is 2) e.g. a 1+ reaction in the

1:8 well is reported as a titre of 8. If the titration end point exceeds 1:2048, testing must be continued by

manual technique with additional dilutions included. Two negative reactions in subsequent dilutions must

follow the end point.

Where possible each sample should be tested in parallel with the previous sample and the results

compared to identify a significant rise in antibody concentration. A rise of at least 2 dilutions is

considered to be a significant increase in titre.

Anti-K antibodies are clinically significant regardless of titre, or reaction strength. Anti-K is the most likely

to cause HDFN other than anti-D and anti-c. While most antibodies associated with HDFN result is

immune destruction of the red blood cells, Anti-K is of greater clinicial significance in HDFN due to the

suppression of erythropoiesis as well as immune red blood cell destruction.






Anti-D and Anti-c titres sample require an aliquot of plasma (minimum of 1 mL of separated plasma) to

be sent to ARCL for an antibody quantitation. If the sample is insufficient for both an antibody titre and

antibody quantitation send the sample for antibody quantition. Refer to local laboratory procedures for

referring samples to ARCL

6.2. Critical Alert Values or Notifications required5,6

If there is a significant increase in the titre end point from the previous titre result the Clinician may be notified refer to local procedure.






6.3. In-House IVD class 1-3 or other Disclaimer Required

Not Applicable

6.4. Record Retention

Refer to NPAAC Retention of laboratory records7.

7. Roles and Responsibilities

1. The Supervising Pathologist discharges the supervision responsibilities as set out in the NPAAC

requirements and Supervision Requirements, including documented review of audits, new tests and

equipment, external QAP, internal Quality Control and Quality Assurance9.

2. Laboratory managers are responsible for ensuring staff are trained in the use of this test method and

are deemed competent. Managers are also responsible for ensuring that records are retained

according to NPAAC requirements7.

3. Technical and Scientific staff are responsible for performing the antibody titration test method and

reporting results as outlined in this document.

8. Legal and Policy Framework

1. Saxo ID Reader II User Manual, DiaMed GmbH Version 2- 08/2016, SAP H007416

2. IH-500 User Manual, DiaMed GmbH Version 2.2- 11/2018, DOC H009225

3. IH-Com user Manual, DiaMed GmbH Version 2-04/2019 DOC H009146

4. NPAAC Requirements for Medical Pathology Services, Third Edition 2018 (Tier 2)

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-medpathserv-

2018

5. NPAAC Requirements for Transfusion Laboratory Practice, Fourth Edition 2019 (Tier 4)

http://www.health.gov.au/internet/main/publishing.nsf/Content/npaac-pub-transfusion

6. ANZSBT Guidelines for Transfusion and Immunohaematology Laboratory Practice, Australasian

Society of Blood Transfusion Inc 1st Edition, revised January 2020 https://anzsbt.org.au/guidelines-

standards/anzsbt-guidelines/

7. NPAAC, Requirements for the Retention of Laboratory Records and Diagnostic Material, Eighth

Edition 2021 (Tier 3) https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-

docs-RetLabRecDI-2018

8. NPAAC, Requirements for Supervision in the Clinical Governance of Medical Pathology

Laboratories, Sixth Edition 2021

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-supervision.htm

9. NSW Health Pathology Supervision Policy NSWHP_PD_019

8.1. Related Documents

Bio-Rad Saxo ID-Reader II NSWHP_PR_057

Bio-Rad IH-500 Immunohaematology Analyser NSWHP_PR_055


https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-medpathserv-2018

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-medpathserv-2018

http://www.health.gov.au/internet/main/publishing.nsf/Content/npaac-pub-transfusion

https://anzsbt.org.au/guidelines-standards/anzsbt-guidelines/

https://anzsbt.org.au/guidelines-standards/anzsbt-guidelines/

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-RetLabRecDI-2018

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-RetLabRecDI-2018

https://www1.health.gov.au/internet/main/publishing.nsf/Content/health-npaac-docs-supervision.htm







Bio-Rad IH-Com Data Management Software NSWHP_PR_054

NSWHP Transfusion Test Methods

9. Review

This procedure will be reviewed by 14/10/2023.

10. Risk

Risk Statement A patient with a high titre antibody can be an indication of HDFN,

or ABMR following an incompatible ABO organ transplant, if

results are not reported accurately, appropriate treatment may not

be given and if this occurs the consequences could result in major

harm to the patient.

Risk Category Clinical Care and Patient Safety

10.1. Key WHS Information

Key WHS Factors and Risks WHS Controls

Reagents and patient

specimens should be treated

as potentially infectious.

Wear PPE while handling or if a spill occurs. Refer to spill clean-

up instructions and product safety data sheets for further details.

11. Further Information

For further information, please contact:

Procedure Contact Officer Position: Chair, Transfusion Clinical Stream

Name: Associate Professor Mark Dean

Telephone: (02) 4320 3894

Email: [email protected]

12. Version History

The approval and amendment history for this document must be listed in the following table.

Version No

Effective Date

Approved By

Approval Date

Procedure Author

Risk Rating Sections Modified

V1.0 27/10/2021 Director Clinical Operations

27/10/2021 Associate Professor Mark Dean

High New Test Procedure



Antibody Titration Test Method - NSW Health Pathology

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