Post on 04-Apr-2018
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DIAGNOSTIC DE LABORATOR
IN HEPATITA CU V.H.B
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ALGORITM DE DIAGNOSTIC IN
HEPATITE
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 (anti)
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Tablou clinic
Icter sclerotegumentar
Astenie fizica
Apetit scazut Febra
Diaree
Urini hipercrome
Scaune decolorate
Nu toate persoanele au tablou clinic manifest.
Adultii si tinerii au manifestari clinice mai frecvent decat copiii.
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AInfectious
Serum
Viralhepatitis
Entericallytransmitted
Parenterallytransmitted
other
E
NANB
B
D
C
HEPATITE VIRALE
PERSPECTIVA ISTORICA
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Source of
virus
Feces
25d (15-50)
blood/
blood-derivedbody fluids
blood/
blood-derivedbody fluids
blood/
blood-derivedbody fluids
Feces
40d (15-60)
Route oftransmission
fecal-oralpercutaneous
permucosal
percutaneouspermucosal
percutaneous
permucosal
fecal-oral
Chronicinfection no yes yes yes no
Prevention pre/post-exposure
Immunization
pre/post-exposure
Immunization
blood donorscreening;
risk behaviormodification
pre/post-exposure
immunization;risk behavior
modification
ensure safedrinking
water
Viral Hepatitis - An Overview
Type of Hepatitis
A B C D E
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Diagnostic V.H.B.
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Structura VHB
ADN/VHB
AgHBc (antigen de centru)
AgHBe(marker deinfectivitate)
AgHBs (antigen desuprafata)
clivare
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Sisteme antigenice VHB
Ag HBs Ac antiHBS
Ag HBc Ac antiHBc
Ag HBe Ac antiHBe
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ETAPE
1. Incubare ser pacient 1h la 37C2. Spalare
3. Incubare conjugat 30min la 37C4. Spalare5. Incubare substrat 20 min la t.camerei6. Citire la spectrofotometru
ELISA de detectie a Ag virale
Ag HBs
Anti HBs
Anti HBs cuplatcu o enzima =conjugat
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Stadii ale infectieio cu V.H.B
Acute Hepatitis B
Chronic Persistent Hepatitis (AgHBe negativ)
Chronic Active Hepatitis (Ag HBe pozitiv)
> Cirrhosis> Hepatocellular Carcinoma
{
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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
ALGORITHM OF LABORATORY DIAGNOSIS
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ALGORITHM OF LABORATORY DIAGNOSIS
HBsAg-
ELISA
Positive=contactwith HBV
negativeAnti-HBc IgM
Positive=
acute
hepatitis B
Negative = must
detect Anti-HBcIgG
Negative =posible
HBsAg
carrier,investigate
Positive=
chronichepatitis B must
detect HBeAg
Positive=
chronic active
hepatitis Negative= chronicpersistent hepatitis
positive=
acutehepatitis B
Negative=nohepatitis B
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Diagnostic molecular in
infectia V.H.B
AMPLIFICAREA TINTEI- PCR
AMPLIFICAREA SEMNALULUI- B-DNA (branched DNA)
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PCRMetoda utilizata pentru detectia ADN-ului de orice tip.
PCR A lifi T t d
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PCR Amplifies a Targeted
SequenceTarget Sequence
DNA Strand
Double Helix
DNA StrandSupercoiled
DNA Strand
Chromosome
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PCR C l St 1
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PCR Cycle - Step 1 -
Denaturation by Heat
Target Sequence
Target Sequence
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PCR Cycle - Step 2 - Primer Pair
Anneals to Ends of Target Sequence
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End of the 1st PCR Cycle - Results in Two
Copies of Target Sequence
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Target Amplification
No. of No. AmpliconCycles Copies of
Target
1 2
2 4
3 8
4 16
5 32
6 64
20 1,048,576
30 1,073,741,82
1 cycle = 2 Amplicon
2 cycle = 4 Amplicon
3 cycle = 8 Amplicon
4 cycle = 16 Amplicon
5 cycle = 32 Amplicon
6 cycle = 64 Amplicon
7 cycle = 128 Amplicon
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Monitorizarea evolutiei naturale /subtratament a hepatitei virale B
Riscul de ciroza creste cu nivelul iv, atat pt persoanele
AgHBe negative, cat si pt cele AgHBe pozitive
Initiere tratamentului pt hepatita cronica BLa persoanele AgHBe negative, iv >10.000 copii/ml indica
infectie activa (marker de infectivitate virala). Tratamentul
este initiat pe baza
PBH iv
Transaminaze serice
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Virus si test Riscrezidual
prin triaj
serologic
Risc rezidual
prin testare
acizi nucleici
Reducere
a riscului
Cost
Per test
(USD)
VHB - ADN/PCR 15.83 9.12 42.4% 30
VHC
ARN/Rt- PCR9.7 2.72 72.0% 200
HIV - Ag p24
2.03
1.48 27.3% 5
HIV - ADN/PCR 1.48 27.3% 50HIV
ARN/Rt-PCR1.01 50.0% 200
Reducerea riscului rezidual asociat transfuziilor