Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The GambiaReportar como inadecuado

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Virology Journal

, 3:23

First Online: 04 April 2006Received: 01 November 2005Accepted: 04 April 2006


Background-AimThe study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.

MethodQuantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.

ResultsThe qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range 1.5 × 10 to 1.5 × 10 copies-mL and correlated well with those from a commercial assay r = 0.91, p < 0.001. Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% 161-209 of HBeAg negative and in all HBeAg positive carriers.

ConclusionThis method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

Electronic supplementary materialThe online version of this article doi:10.1186-1743-422X-3-23 contains supplementary material, which is available to authorized users.

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Autor: Maimuna E Mendy - Steve Kaye - Marianne van der Sande - Pura Rayco-Solon - Pauline A Waight - Deborah Shipton - Dorka Awi


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