A real-time quantitative assay for hepatitis B DNA virus (HBV) developed to detect all HBV genotypes

Abstract

SUMMARYHepatitis B virus (HBV) is a major cause of chronic liver disease worldwide. Besides genotype, quantitative analysis of HBV infection is extensively used for monitoring disease progression and treatment. Affordable viral load monitoring is desirable in resource-limited settings and it has been already shown to be useful in developing countries for other viruses such as Hepatitis C virus (HCV) and HIV. In this paper, we describe the validation of a real-time PCR assay for HBV DNA quantification with TaqMan chemistry and MGB probes. Primers and probes were designed using an alignment of sequences from all HBV genotypes in order to equally amplify all of them. The assay is internally controlled and was standardized with an international HBV panel. Its efficacy was evaluated comparing the results with two other methods: Versant HBV DNA Assay 3.0 (bDNA, Siemens, NY, USA) and another real-time PCR from a reference laboratory. Intra-assay and inter-assay reproducibilities were determined and the mean of CV values obtained were 0.12 and 0.09, respectively. The assay was validated with a broad dynamic range and is efficient for amplifying all HBV genotypes, providing a good option to quantify HBV DNA as a routine procedure, with a cheap and reliable protocol.

Comparison of a novel real-time PCR method (RTA) and Artus RG for the quantification of HBV DNA and HCV RNA

“…However, some steps of the protocols used require intensive effort and they have some risks. RT-PCR is used as a reasonably standardized diagnostic tool for a large number of pathogenic viruses, and substitutes other virological diagnostic methods because of its high specificity and wide dynamic range [8]. It is recommended to use standardized methods in clinical practice for the efficient monitoring and control of HBV infections.…”

A novel colorimetric PCR-based biosensor for detection and quantification of hepatitis B virus

“…Three different hairpin probes (probe-HBV-1, probe-HBV-2and probe-HBV-3) and corresponding primers were designed against the different conserved regions of the HBV genome including overlapping genes encoding X-protein, S gene and X gene region [2,21,22] (Table S2, S3 & S4). Under the optimized PCR amplification conditions of corresponding probes, the different colorimetric results by using those three pairs of probes were turned out (Fig.…”

Occult hepatitis B viral infection among blood donors in South–Eastern Nigeria

“…2,4 With qPCR, using TaqMan chemistry, ultra-sensitivity, reliability and quantitation of viral load are achieved rapidly at lower cost. 26 …”