Tes Lab Pada Peny Infeksi Dan Tropis (Lebih Singkat)

8/18/2019 Tes Lab Pada Peny Infeksi Dan Tropis (Lebih Singkat)

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Bagian Patologi Klinik

FK UNHAS



Pengamatan Pada :

- Eritrosit, Lekosit, Trombosit

- Maniestasi : Anemia, lekositosis !leko"eni dan #$%

Lekositosis- Um&mn'a Netroil , bent&k m&da

- Netroilia lan(&t ineksi kronik

- Netroilia meng)ebat * sel m&da reaksi le&kemoid

+ Non-anas ./-01 2 314!5l

+ $nlamasi, Stress, Tra&ma6



Anemia

+ Bisa timb&l sekali"&n 7adangan besi 7&k&"

+ Anemia akut : "erdara)an ! destr&ksi eritrosit9misaln'a : 7old agl&tinin se)&b&ngan dengan

M'7o"lasma "ne&moniae

+ Anemia kronik, dengan :

- %adangan besi 'ang normal ata& meninggidi sistem retik&loendotelial

- Pen&r&nan besi dalam "lasma

- Pen&r&nan T$B%



$neksi seri&s * bakteremia

+ ram negati #$% 9gram "ositi k&rang

- Tromb&s

- PT meman(ang- F#P

- Fibrinogen

Trombosito"enia bisa (&ga men(adi tanda se"sis

bakterial dan bisa bermanaat dalammengobser;asi res"on "asien ter)ada" tera"i6



Laborator' indings

+ Hematolog'- Le&ko"enia

- Trombo7'to"enia

- ser&m aminotranserase 9AST, ALT ele;ations

+ T)e diagnosis is made b' lab test seroloimm&nolog'

36 Hemagl&tination tests

.6 %om"lement Fi2ation test

06 Ne&trali<ation Test

- $gM EL$SA or

- "aired serolog' d&ring re7o;er' or

- b' antigen-dete7tion EL$SA or

- =T-P%= d&ring t)e a7&te ")ase

+ 8ir&s is readil' isolated rom blood in t)e a7&te ")ase i mos>&ito

ino7&lation or mos>&ito 7ell 7&lt&re



Diagnosis Demam Dengue/Demam BerdarahDengue



S&s"i7ion o deng&e ine7tion

#ela' ater onset o e;er

Unkno?n or @ / da's / )ari

NS3 Antigen Antisi"ated #iagnosis $gM!$g Serologis $m"ro;e "atient Management

* - * -

%onirmed earl' $gM serolog' Late a7&te or #eng&e

ine7tion

A7&te ine7tion "ast-ine7tion is&nlikel'

* -

Presumably early early acute infection

Acute infection is unlikely A se7ond sam"le isre>&ested or 7onirmation

EA=L #$ANS$S LATE #$ANS$S



$n deng&ePresent b' t)e .nd da' o e;erB' t)e Ct) or /t) da', t)e ?b7 7o&nt .111 to C111!ml,

.1 to C1 D gran&lo7'tesModerate alb&min&ria and a e? 7asts ma' be o&nd

- #eng&e ma' be 7on&sed ?it) %olorado ti7k e;er,

t'")&s, 'ello? e;er, or ot)er )emor)agi7 e;ers6 Serologi7 diagnosis ma' be made b'

- Hemaggl&tination in)ibiting and 7om"lement i2ation test &sing "aired sera

- b&t is 7om"li7ated b' 7ross-rea7tions ?it) ot)er

la;i;ir&s antibodies



$n deng&e )emorr)agi7 e;er• H7t /1 D: i"resent d&ring s)o7k

• B% 7o&nt in 3!0 o "atients

• %oag&lai;e abnormalities

- Trombo7'to"enia 93116111!ml

- "ositi;e to&rni>&et test- "rolonged PT

- Minimal "rotein&ria ma' be "resent6

- AST le;els ma' be moderatel'

- Serologi7 test &s&all' s)o? )ig) 7om"lement

i2ation antibodies titers againts la;i;ir&s,

s&ggesti;e o a se7ondar' imm&ne res"onse



H 7lini7al 7riteria or diagnosis o deng&e

)emorr)agi7 e;er :• A7&te onset o )ig), 7ontino&s e;er lasts or . to G

da's• Hemorr)agi7 maniestations, in7l&ding at least a

"ositi;e to&rni>&et tes and "ete7)iae, "&r"&ra,e77)'moses, bleeding g&ms, )ematemesis or melena• He"atomegal'• Trombo7'to"enia 93116111!mlor )emo7on7entration

9H7t in7reased b' .1D• T)ose ?it) deng&e s)o7k s'ndrome also )a;e a ra"id

?eak "&lse ?it) narro?ing o t)e "&lse "ress&re 9.1mmHg or )'"otension ?it) 7old, 7lamm' skin andrestlessness6



Laborator' tests are generall' not ne7essar'

9;iral 7&lt&res and T<an7k smear ?ill 7onirmdiagnosis in "atient ?it) at'"i7al "resentation

Antibod' to a""ro"riate serot'"e :- Sero7on;ersion- $n7rease- #ire7t imm&nol&ores7ent antibodies slide test9ra"id diagnosis

T<an7k "re"aration :

- Base o lesion

- M&ltin&7leate giant 7ells



Laborator' test are generall' notne7essar' 9;iral 7&lt&res andT<an7k smear ?ill 7onirm

diagnosis in "atient ?it) at'"i7al"resentation

T)e T<an7k "re"aration s)o?s

m&ltin&7leate giant 7ells or bot);ari7ella-<oster ;ir&s and HS8



#ara)

- leko"eni

- ser&m amilase dalam 31 )ari

- Serologi :

+ 7old aggl&tinin+ $gM , ma2 . mingg&, meneta" 4-I b&lan

kadar ser&m kon;alesens C2 dr "d ser&m ak&t

+ tes i2asi kom"lement t)d atb "ositi mingg&

"ertama

Biakan

- 8ir&s dari l&da) 3-/ )ari



Kom"likasi :

- $nlamasi testis ! o;ari&m : lekositosis, LE#

- Pankreatitis : lekositosis, amilase , )i"erglikemia

- Meningitis : sel L%S /11!5l, monon&klear gl&kosa

normal, "rotein agak 9.1-3./ mg!dl



Tem&an Laboratori&m

- #ara) :

+ Lekosit ter&tama limosit dan segmen

+ Serologi : E$A

- $gM : Fase ak&t 9 J 3-. )ari- $g : 31 )ari

- Sekret :

+ A"&san * "&lasan im&nol&orosen

+ P&lasan T<an7k : M&ltin&7leated iant %ells

- Biakan :

+ Ba)an : sekret res" dan &rin

+ $dentiikasi : (aringan



Tes Lab 'ang bisa dilak&kan:

- Sediaan A"&s :

+ Ba)an : Kerokan dasar ;esikel

+ P&lasan : T<an7k M&ltin&7leated iant %ells

+ Sensiti;itas J 41 D

- #ara) :+ Serologi :

- Titer atb ser&m kon;alesent C2 d"d

ser&m ak&t6

- Hemagl&tinasi

- EL$SA

- FAMA

+ P%= : deteksi #NA 8ir&s



#NA t'"ing : %ir7&lar do&bel stranded ,111 b"

+ %ross)'bridi<ation :- /1D : t'"e se"eration

- /1D : s&bt'"e se"eration



enerall' not ne7essar'

ram stain and %S to 7onirm t)e diagnosis?)en t)e 7lini7al "resentation is &n7lear

Sedimentation rate "arallel to a7ti;it' o t)edisease

Anti-#NAse B and ant' )'al&ronidase

Urinal'sis : )emat&ria ?it) er'tro7'te 7asts and

"rotein&ria in "atients ?it) a7&te ne")ritis



LABORATORY DIAGNOSIS (ANTIBODY TEST)

1. Enzyme-linked Immunoso!en" Ass#y (ELISA)

$os" %ommonly used !e%#use o& i"s el#"i'ely simple me"odoloy* high sensitivity #nd sui"#!ili"y &o

"es"in large numbers of samples* +#"i%ul#ly in !lood"es"in %en"es.

Simul"#neously de"e%"ion o& #n"i!odies "o HIV-1 andHIV-2 in um#n seum o +l#sm#.

De"e%" #n"i!odies die%"ed ##ins" m#,o ou+ o& I-1#nd I-/ +o"eins0

I-10 en'elo+e +o"ein o& +1%oe s"u%"u#l +o"ein o& +/

I-/0 en'elo+e +o"ein o& +23

$e"ods0 indie%"* %om+e"i"i'e* #n"ien s#nd4i%



ELISA &o I #n"i!ody

$i%o+l#"e ELISA &o I #n"i!ody0 %oloued 4ellsindi%#"e e#%"i'i"y



ELISA Tes" 5i" ELISA $#%ine

LABORATORY DIAGNOSIS (ANTIBODY TEST0 ELISA)



De"e%"ion o& A 6 A! in # sinle "es"

7"ili"y in +im#y in&e%"ion0+e-seo%on'esion (84indo4 +eiod9)

Blood !#nk

Au"om#"ed +l#"&oms #'#il#!le

LABORATORY DIAGNOSIS (ANTIBODY TEST0 ELISA)



/. Sim+le Tes"

Re:uies e#"e "#n 2; minu"es !u" #s +o%edues "#"%#n !e +e&omed e#sily without instrumentation.

<i"in "is %l#ss o& "es"s #e agglutination assays in 4i%

#n"ien-%o#"ed +#"i%les (ed !lood %ells =RB>?* l#"e@+#"i%les o el#"in +#"i%les) #e #llo4 "o e#%" 4i" seum#n"i!odies "o &om 'isi!le %lum+in (#lu"in#"ion).

I& RB> #e used* "e "e%ni:ue is "emed +#ssi'e

#em#lu"in#"ion (A) l#"e@ +#"i%les is kno4n #s l#"e@#lu"in#"ion (LA) el#"in +#"i%le is kno4n #s +#"i%le#lu"in#"ion (A).



Good sensitivity, low cost #nd ease of performance.

I" in%o+o#"es # uality system to detect nonspecificantibodies die%"ed "o4#d "e el#"in +#"i%les "emsel'ess#m+le need "o !e "e#"ed !y #dso+"ion #nd e"es"ed.

C#lse-ne#"i'e e#%"ion due "o pro!one (ini!i"ion o&#lu"in#"ion 4en e@%ess o& #n"i!ody +esen"* e'en"s

o+"im#l %om!in#"ion) - +e&om dilu"ion.

"esults %#n !e o!"#ined within 2 hours.



#$%&"$'&"( )I$G*&+I+ $*'I%&)( '+' +I/0# '+'



#"i%le Alu"in#"ion Tes"



(B) Su++lemen"#y>on&im#"ion Tes"s

1. estern %lot %

Te mos" 4idely #%%e+"ed 3gold standard4 &o '#lid#"ion o&I esul"s.

B#sed on ele%"o+oe"i% "e%ni:ue "o se+##"e I #n"iens

dei'ed &om viral lysate grown in culture #nd "e se+##"e#n"iens #e "#ns&eed (!lo""ed) on"o ni"o%ellulosemem!#ne.

5ommercial % #e su++lied 4i" indi'idu#l s"i+s o&ni"o%ellulose %on"#inin "e !lo""ed* se+##"ed I #n"iens.A++li%#"ion o& #I+$ techniue in de"e%"ion o& s+e%i&i%#n"i!odies "o e#% 'i#l #n"iens.

/odified % #s "e #!ili"y "o identify and differentiate in&e%"ions !y HIV-1 and HIV-2.



<es"en Blo"

LABORATORY DIAGNOSIS (ANTIBODY TEST0 S7LE$ENTARY>ONCIR$ATORY)



2. #ine Immunoassay #I$

Ano"e #l"en#"i'e "o "e %l#ssi% <es"en !lo".

"ecombinant or synthetic peptide antigens #e #++lied on"onylon s"i+ #"e "#n ele%"o+oesed #s in "e <es"en

!lo".

7se o& #"i&i%i#lF #n"iens decreases the presence ofcontaminating substances dei'ed &om %ell %ul"ue "#" %#n%#use in"e&een%e #nd some"imes false reactions.

A num!e o& e+o"s #'e 'ei&ied "#" "e accuracy iseuivalent to the estern blot.






Line Immuno#ss#y



In"e+e"#"ion %i"ei# &o Immuno!lo" "es"s

O#niz#"ion $inimum !#nd e:uiemen"s &oe#%"i'eF +#""en

Amei%#n Red >oss A" le#s" one band from each gene +odu%" ou+0 # AND +ol AND en'

>en"es &o Dise#se $ny two o& +/* +1 o +1/;13;>on"ol (>D>)

<old e#l" O#niz#"ion 'wo env bands (+13;* +1/; o +1)(<O) - +ol !#nds* - # !#nds

Du on" p26 AND p71 AND gp61 or gp12891:8



gp160

gp120

p68

p55p53

gp41-45

p40

p34

p24

p18

p12

gp160

gp120

p68

p55p53

gp41-45

p40

p34

p24

p18

p12

gp160

gp120

p68

p55p53

gp41-45

p40

p34

p24

p18

p12

earl' re7ent ! establis)ed ad;an7ed

S+e%"umS+e%"umo& #n"i-Io& #n"i-I

"es"in"es"in

DNA PCRDNA PCR

RNA PCRRNA PCR

p24 Agp24 Ag

3rd gen ELISA

1st gen ELISA

1wk 2wk 3wk 2m 6m 1!r 2!r 3!r "8!r



I RAID TEST 5IT



A 4om#n demons"#"es "e use o& "e I #+id "es"

LABORATORY DIAGNOSIS (ANTIBODY TEST0 RAID TEST)

http://en.wikipedia.org/wiki/Image:Oraquick.jpg



L B R RY D GN ( N B DY E )




I Tes"in S"#"eies

<O e%ommends "ee "es"in s"#"eies "o0

-/a<imi!e accuracy

-/inimi!e cost

>oi%e o& s"#"ey de+ends u+on0

-ob=ective o& "es"

-prevalence of HIV in +o+ul#"ion

LABORATORY DIAGNOSIS (ANTIBODY TEST0 TESTING STRATEGIES)



( )

&b=ective of testing 0revalence

of Infection

'esting

+trategyT#ns&usiondon#"ion

s#&e"y

All I

Su'eill#n%eK1; I

M1; II

Di#nosis

>lini%#l sins

sym+"oms o& Iin&e%"ionAIDS

#ll II

Asym+"om#"i%

K1; II

M1; III

<O Tes"in S"#"eies



OSITIEi Risk Gou+ (<O /-"es"s s"#"ey)

ELISA - Re#%"i'eA - De"e%"ed

Lo4 Risk Gou+ (<O 2-"es"s s"#"ey)ELISA - Re#%"i'eA - De"e%"edImmuno!lo" - osi"i'e

NEGATIEBo" s%eenin "es"s0 EIA - non-e#%"i'e #nd

A - no"-de"e%"ed



LABORATORY DIAGNOSIS (ANTIBODY TEST0 INTERRETING TEST RES7LTS )

CALSE OSITIE

Te ELISA "es" m#y +odu%e &#lse +osi"i'e esul"s on some!lood &om unin&e%"ed indi'idu#ls.

Em+loys #" le#s" 2 different tests.

$#y due "o nons+e%i&i% e#%"ion* #u"oimmune dise#ses(Sys"emi% Lu+us Ey"em#"osus)* e"%



CALSE NEGATIE

Tes" %on%ludes I is no" +esen"* 4en in &#%" "e +eson isin&e%"ed

Tis o%%us 4en "e !lood is "#ken duin "e windowperiod.

I& "ee #s !een e@+osue "o isk #%"i'i"ies* i" is #d'is#!le"o e+e#" "e "es" 2-3 mon"s l#"e.

LABORATORY DIAGNOSIS (ANTIBODY TEST0 INTERRETING TEST RES7LTS )



Sediaan darah malaria

Kegunaan sediaan darah malaria :

Menentukan ada tdknya parasit malaria

Menentukan spesies & stadium plasmodium

Dapat melacak ! " !! parasit#$L darah

Menentukan kepadatan parasit

/1



Sediaan darah tebal

%ara terbaik menemukan parasit malaria

Mudah dibuat

Diperiksa paling sedikit !! lapangan pandang

Sediaan darah tipis

Digunakan utk identifikasi enis Plasmodium biladgn sediaan darah tebal sulit ditentukan

Diperiksa paling sedikit '!! lapangan pandang

/3



Kelebihan sediaan darah tebal

Lebih banyak sel darah yg diperiksaParasit lebih mudah ditemukan

Kekurangan sediaan darah tebal

(dk dpt membandingkan ukuran Plasmodium dgn

ukuran eritrosit

Spesies Plasmodium sukar ditentukan

/.



,entuk (ropo-oit%incin kecil .⅓ ө eritrosit normal/

Sitoplasma biru

Kromatin inti merah

,entuk Ski-on

*arang ada dlm sirkulasi darah tepi

*k ditemukan dlm darah tepi tandamalaria berat

/C



,entuk 0ametosit

Sgt khas yaitu elips .crescent/

,erpigmen 1arna hitam

Sitoplasma kuning

//



(eknik Quantitative Buffy Coat .2,%/

Prinsip : (es 3luoresensi

Eritrosit yg terinfeksi Plasmodium akan terlihat

berfluoresensi di b1h mikroskop fluoresensi%epat namun peralatannya mahal

(dk dpt membedakan spesiesPlasmodium & tdk dpt digunakan utk

hitung parasit+

/4



Mendeteksi Ab#Ag spesifik terhadap parasitmalaria atau eritrosit yg terinfeksi Plasmodium

(es imunoserologis yg melacak Ab tdk dipakai

utk keperluan diagnosis(es imunoserologis malaria :

Radioimmunoassay .45A/

Enzyme Linked Immunoassay .EL5SA/

Immunochromatographi .5%(/

Indirect Fluorescent Antiody !est .53A(/

/G



/

4adioisotop sebagai label

Kadar Ag atau Ab pada sampel dpt ditentukan scr

kuantitatif Lo" detection limit 6! parasit#$L darah

Sensitif

Kurang praktis & berbahaya



/I

Lebih praktis dibanding 45A

En-yme direaksikan dgn substrat kromogenintensitas 1arna sebanding dgn kadar bahan

Mendeteksi Ag & Ab spesifik terhadap Plasmodium



41

Rapid #iagnostic !est

Melacak Ag parasit malaria melalui pengikatan

Ag oleh Ab monoklonalAda 7 enis Ag utama yg sering diadikantarget 5%( utk mendiagnosis malaria yaitu :

$istidine%rich protein .84P/

&arasite specific lactate dehydrogenase .pLD8/

Plasmodium Aldolase



43

Keunggulan tes 5%(

Praktis

(dk membutuhkan alat pembantu lain(dk memerlukan tenaga terampil

Kelemahan tes 5%(

8anya dpt melacak parasit 9 !! parasit#$L darah

Membutuhkan biaya pemeriksaan yg relatif sdg

(dk dpt memberi informasi deraat parasitemia



4.

Mendeteksi Ab spesifik terhdp Plasmodium

Keadaan dimana parasit sangat minimal

(dk utk menentukan infeksi baru

Mendeteksi keempat spesies PlasmodiumManfaat utk penelitian epidemiologi



Mendeteksi DA spesifik terhadap parasitPlasmodium dlm darah penderita malaria

(eknik &olymerase chain reaction .P%4/

Dpt melacak sampai 6 parasit#$L drh

Dpt mengidentifikasi spesies Plasmodium

;aktu pemeriksaan cepat

Sensiti<itas & spesifisitasnya tinggi

Mahal

40



enis tes laboratori&m &nt&k t&berk&losisterdiri dari: Tes Mikrobiologi, terdiri dari:

Tes sel&ler:

Tes BTA S"&t&m Tes Biakan dan $dentiikasi M.tuberculosis

Tes Ke"ekaan Antibiotika

Tes molek&ler: P%=

Tes Serologis, terdiri dari: Semira"id: TB-#ot, EL$SA, Tb-kom"leks

=a"id: M'7odot, $%T-TB



Anamnesis

Pemeriksaan Fisis Pemeriksaan =adiologis

Tes Laboratori&m

.4 Maret 1GU(ian Ak)ir Magister!U(ian Lokal 4/

Tes MikrobiologiTes sel&ler: Tes A"&san BTA

Tes Biakan $dentiikasi

Tes Ke"ekaan Antibiotika

Tes molek&ler: P%=Tes Serologis

Semira"id: TB-#ot, EL$SA, Tb-kom"leks

=a"id: M'7odot, $%T-TB se"erti M'7ote7 TB



Mikroskopik =iehl eelsen

Dekontaminasi

Kultur .pembiakan/ medi&m Lo?enstein-ensen

P%4

5solasi DA: Metode ,oom Proses : denaturasi> annealing> elongasi

5S?!

.4 Maret 1GU(ian Ak)ir Magister!U(ian Lokal 44



(es 5munoserologi #eteksi Ab ter)ada" Ag mikobakterial s"esiik

ata& gab&ngan bebera"a antigen Ag41: EL$SA

Ag34 : 34 k#a,

s"esies-s"esii7 e"ito" im&nodominan

stadi&m a?al ineksi M6tb7 TB "rimer 0k#a

s"esies-s"esii7 e"ito" im&nodominan

ESAT-4

kontak bar& ter(adi, kon;ersi meningkatn'a risiko"en'akit

.4 Maret 1GU(ian Ak)ir Magister!U(ian Lokal 4G



K&lt&r :

+ #ara) : "ositi dalam 31 )ari "ertama

+ Tin(a &rin "ositi dalam mingg& 0-/

+ S&ms&m t&lang Serologi :

+ Tes idal : Ser&m semb&) C2 dr"ada sakit

#ara) r&tin : leko"eni



Feli2-idal test

- meas&res le;els o aggl&tinating antibodies against and Hantigens

- &s&all' a""ear ater /- da's o ine7tion - limited 7lini7al &tilit' in a7&tel' ill "atients be7a&se "ositi;e res&lts

ma' re"resent "re;io&s ine7tion in endemi7 areas

- "aired titration s)o&ld be "reormed

- $n t)e absen7e o re7ent imm&ni<ation, a )ig) titre o antibod' to

antigen 3:4C1 is s&ggesti;e b&t not s"e7ii76- alse "ositi;e 97ross rea7ti;it'

An EL$SA or antibodies to t)e 7a"s&lar "ol'sa77)aride 8i antigen is&se&l or dete7tion o 7arriers b&t not or t)e diagnosis o a7&teillness



Typhidot test that detects presence of IgM and IgG in one hour(sensitivity>95%, Specificity 75%)

Typhidot-M, that detects IgM ony (sensitivity 9!% andspecificity 9"%)

Typhidot rapid (sensitivity #5% and Specificity 99%) is arapid $5 inute iunochroatographic test to detect IgM&

IgM dipstick test

Tes Lab Pada Peny Infeksi Dan Tropis (Lebih Singkat)

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Transcript of Tes Lab Pada Peny Infeksi Dan Tropis (Lebih Singkat)