Real-time PCR assays for hepatitis B virus DNA quantification may require two different targets

Virology Journal
Chao LiuLunan Wang

Abstract

Quantification Hepatitis B virus (HBV) DNA plays a critical role in the management of chronic HBV infections. However, HBV is a DNA virus with high levels of genetic variation, and drug-resistant mutations have emerged with the use of antiviral drugs. If a mutation caused a sequence mismatched in the primer or probe of a commercial DNA quantification kit, this would lead to an underestimation of the viral load of the sample. The aim of this study was to determine whether commercial kits, which use only one pair of primers and a single probe, accurately quantify the HBV DNA levels and to develop an improved duplex real-time PCR assay. We developed a new duplex real-time PCR assay that used two pairs of primers and two probes based on the conserved S and C regions of the HBV genome. We performed HBV DNA quantitative detection of HBV samples and compared the results of our duplex real-time PCR assays with the COBAS TaqMan HBV Test version 2 and Daan real-time PCR assays. The target region of the discordant sample was amplified, sequenced, and validated using plasmid. The results of the duplex real-time PCR were in good accordance with the commercial COBAS TaqMan HBV Test version 2 and Daan real-time PCR assays. We showed that two ...Continue Reading

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Citations

Sep 29, 2018·Genes·Emilia Hadziyannis, Andreas Laras
Aug 8, 2019·Indian Journal of Hematology & Blood Transfusion : an Official Journal of Indian Society of Hematology and Blood Transfusion·Chao LiuLunan Wang
Aug 4, 2020·Royal Society Open Science·Kashif Aziz Khan, Peter Cheung
May 11, 2020·Biotechnology and Applied Biochemistry·Ting-Ting YiYan Cai

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Methods Mentioned

BETA
PCR

Software Mentioned

SPSS
DNAman
Graph Prism

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