Renfei Lu1, Jianguo Shao1, Min Zhou2, Hongping Zhang2, Yueping Wu1. 1. Department of Laboratory, The Third People's Hospital of Nantong, Nantong, Jiangsu, P. R., China. 2. Nantong Center for Disease Control and Prevention, Nantong, Jiangsu, P. R., China.
Abstract
BACKGROUND: Hepatitis B virus (HBV) G1896A mutation was associated with HBeAg seronegativity and hepatitis B related acute-on-chronic liver failure. In this study, we developed Taqman amplification refractory mutation system (Taqman-ARMS) and established a strict control system to detect HBV G1896A mutant. METHODS: HBV viral DNA was isolated from 60 patient serum samples, and full-length HBV genome was cloned. Then, Taqman-ARMS was used to detect HBV G1896A mutant. RESULTS: The assay has the sensitivity of 1E+3 IU/ml G1896A template, and 0.1% weak population virus with G1896A could be found in mixtures. Total of all 60 clinical samples random collected were detected by Taqman-ARMS, the results were consistent with those by DNA sequencing. CONCLUSION: The proposed Taqman-ARMS real-time PCR method for the detection of G1896A mutation of HBV was rapid, simple, sensitive, specific, and applicable in the clinical setting.
BACKGROUND:Hepatitis B virus (HBV) G1896A mutation was associated with HBeAg seronegativity and hepatitis B related acute-on-chronic liver failure. In this study, we developed Taqman amplification refractory mutation system (Taqman-ARMS) and established a strict control system to detect HBVG1896A mutant. METHODS:HBV viral DNA was isolated from 60 patient serum samples, and full-length HBV genome was cloned. Then, Taqman-ARMS was used to detect HBVG1896A mutant. RESULTS: The assay has the sensitivity of 1E+3 IU/ml G1896A template, and 0.1% weak population virus with G1896A could be found in mixtures. Total of all 60 clinical samples random collected were detected by Taqman-ARMS, the results were consistent with those by DNA sequencing. CONCLUSION: The proposed Taqman-ARMS real-time PCR method for the detection of G1896A mutation of HBV was rapid, simple, sensitive, specific, and applicable in the clinical setting.
Authors: Lei Xiao; Bin Zhou; Hongbo Gao; Shiwu Ma; Guifeng Yang; Min Xu; William G H Abbott; Jinjun Chen; Jian Sun; Zhanhui Wang; Jinlin Hou Journal: J Med Virol Date: 2011-09 Impact factor: 2.327
Authors: W F Carman; M R Jacyna; S Hadziyannis; P Karayiannis; M J McGarvey; A Makris; H C Thomas Journal: Lancet Date: 1989-09-09 Impact factor: 79.321