Comparison of serum hepatitis B virus replication markers in patients with chronic hepatitis B: studies on HBeAg/Anti-HBe system, viral dna polymerase and HBV-DNA Comparação dos marcadores sorológicos da replicação do vírus da hepatite B: estudos sobre o sistema AgHBe/anti-HBe, DNA polimerase virai e HBV-DNA

The detection of HBV-DNA in serum by molecular hybridization is the most sensitive and specific marker of replication and infectivity of hepatitis B virus and currently is proposed as a routine diagnostic technique in the follow-up of HBV - related diseases. Comparing different techniques already de...

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Bibliographic Details
Published in:Revista do Instituto de Medicina Tropical de São Paulo
Main Authors: João Renato Rebello Pinho, Luís Edmundo Pinto da Fonseca, Yu Song, Yuriko Miyamoto, Flair José Carrilho, Celso Francisco Hernandes Granato, Luiz Caetano da Silva
Format: Article in Journal/Newspaper
Language:English
Published: Universidade de São Paulo (USP) 1989
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Online Access:https://doi.org/10.1590/S0036-46651989000500006
https://doaj.org/article/8da92c537e8e4226949ad73007fb376b
Description
Summary:The detection of HBV-DNA in serum by molecular hybridization is the most sensitive and specific marker of replication and infectivity of hepatitis B virus and currently is proposed as a routine diagnostic technique in the follow-up of HBV - related diseases. Comparing different techniques already described, we found that direct spotting of serum samples on nitrocellulose membranes under vacuum filtration, followed by denaturing and neutralizing washes is more practical, simple, sensible and reproducible. DNA polymerase assay using phosphonoformic acid as specific viral inhibitor has shown 86.8% of concordance with HBV-DNA detection, and so, it is an useful alternative in the follow-up of hepatitis B chronic patients. We found 19.2% HBeAg positive samples with no other markers of viral replication and no anti-HBe positive sample had detectable HBV-DNA. Discordance between the 2 systems have been extensively described, and we confirm this for the first time in our country. Molecular biological techniques are essential to determine the replication status of chronic hepatitis B patients. A detecção do genoma do HBV no soro por hibridização molecular é o mais sensível e específico marcador da replicação e infectividade do HBV, sendo sua utilização proposta como técnica rotineira no acompanhamento de doenças relacionadas a este vírus. Comparando diferentes técnicas descritas anteriormente, escolhemos a deposição direta das amostras séricas sobre a membrana de nitrocelulose sob filtração a vácuo, seguida de banhos desnaturantes e neutralisantes como mais prática e simples, com sensibilidade equivalente. O ensaio da DNA polimerase usando ácido fosfonofórmico como inibidor virai específico mostrou 86.8% de concordância com a detecção direta do DNA viral, sendo, portanto, uma alternativa viável no acompanhamento de pacientes com hepatite crônica B. Encontramos 19,2% das amostras AgHBe positivos sem outros marcadores de replicação viral. Por outro lado, nenhuma amostra anti-HBe positiva teve HBV-DNA detectável. ...