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DIAGNOSTIC DE LABORATORIN HEPATITE DE ETIOLOGIE

VIRALA

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ORIENTAREA DIAGNOSTICA INHEPATITE

1. Tablou clinic

2. Teste biochimice Markerii necrozei hepatocitare (transaminaze, LDH)

Markeri colestaza (FAL, GGT, bilirubina)

Markeri ai incapacitatii de sinteza la nivel hepatic (proteine serice,colesterol, trigliceride)

3. Date epidemiologice: cale transmitere, contactinfectant, per. incubatie, vaccinuri

4. Diagnostic etiologic = detectia markerilorvirali:

antigene virale (Ag)

anticorpi antivirali (Ac)

acid nucleic viral

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Diagnostic V.H.B.

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ANTIGENE VHB

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Sisteme antigenice VHB

Ag HBs Ac antiHBS

Ag HBc Ac antiHBc

Ag HBe Ac antiHBe

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Semnificatia markerilor de hepatita B Antigenul HBs, markerul cel mai precoce al infectiei VHB,

persista in cursul hepatitei acute si in prima parte aconvalescentei. Prezenta sa dupa mai mult de 6 luniindica hepatita cronica persistenta sau activa.

Anticorpii anti HBc IgM apar in cursul bolii acute si setransforma apoi in anticorpi anti HBc IgG care persista

indefinit in serul pacientilor indicand o infectie trecuta sauprezenta (semnul de trecere prin boala). Cel mai bun marker serologic de infectivitate este

considerat antigenul HBe este pozitiv in hepatita acutaB, dar si in hepatita cronica agresiva B.

Prezenta anticorpilor anti HBe semnifica infectie VHB incurs de vindecare.

Testarea existentei imunitatii impotriva VHB se realizeazaprin detectia anticorpilor antiHBs (marker de vindecare- inprezenta anticorpilor anti HBc IgG pozitivi sau devaccinare- in absenta anticorpilor anti HBc IgG .

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Hepatita B

Hepatita acutaHepatita cronica persistenta

Hepatita cronica agresiva Ciroza hepatica

Carcinom hepato-celular

ALGORITM

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Simptome

AgHBeADN/VHB

anti-HBe

anti-HBc totali

anti-HBc IgM anti-HBsAgHBs

0 4 8 12 16 20 24 28 32 36 52 100

Infectia acuta cu VHB

evolutie spre vindecare

Saptamini post expunere

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IgM anti-HBc

Total anti-

HBc

HBsAg

Acute(6 months)

HBeAg

Chronic(Years)

anti-HBe

0 4 8 12 16 20 24 28 32 36 52 Years

Weeks after Exposure

Infectia cronica cu VHB

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Tehnica ELISA sandwich(godeuri peliculizate cu anticorpi monoclonali specifici (Mab) impotriva Ag ce urmeaza a

fi detectat)

1. Incubare ser pacient 1h la 37C2. Spalare

3. Incubare CONJUGAT (al 2lea Mab specific impotriva Ag ce urmeaza a fidetectat) 60min la 37C

4. Spalare5. Incubare SUBSTRAT 20 min la t.camerei6. Solutie de stopare7. Citire la spectrofotometru => D.O., MN, MP, C.O.

ELISA de detectie a Ag virale

Ag HBs

Anti HBs

CONJUGAT=Anti HBs cuplatcu o enzima

SUBSTRAT

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Spalator

automat

Distribuirea probelor cupipeta automata

http://www.mtxlsi.com/FinstripWasherSpecs.htmhttp://www.mtxlsi.com/FinstripWasherSpecs.htm

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Spectrofotometru

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Diagnostic V.H.D.

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RNA

Hepatitis D Virus

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anti-HBs

Symptoms

ALTElevated

Total anti-HDV

IgM anti-HDV +IgM anti HBc

HDV RNA

HBsAg

Coinfectia HBV - HDV

Timp dupa expunere

Titru

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Jaundice

Symptoms

ALTanti-HDV+ anti HBc IgG

IgM anti-HDV

HDV RNA

HBsAg

Suprainfectia HBV - HDV

Timp dupa expunere

Titre

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Diagnostic

V.H.A.

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0 1 2 3 4 5 6 7 8 9 10 11 12 13saptamana

simptomatologieALT

IgMIgG

VHA in MFInfectieViremie

EVOLUTIA MARKERI DE INFECTIE VHA

HEPATITA A NU CRONICIZEAZA!!

Diagnostic hepatita acuta A = detectie Ac anti VHA IgM (ELISA de captura)

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ELISA de capturapentru detectia clasei de anticorpi

Anti-IgM

IgM

Ag marcatenzimatic =conjugat

ETAPE

1. Incubare ser pacient 1h la 37C

2. Spalare

3. Incubare conjugat 30 min la 37C

4. Spalare

5. Incubare substrat 20 min la t. camerei

6. Citire la spectrofotometru