Abe A, Inoue K, Tanaka T, Kato J, Kajiyama N, Kawaguchi R, Tanaka S, Yoshiba M, Kohara M
Center for Molecular Biology and Cyotogenesis, SRL, Inc., Hino Tokyo 192-0002, USA.
J Clin Microbiol. 1999 Sep;37(9):2899-903. doi: 10.1128/JCM.37.9.2899-2903.1999.
Quantitation of hepatitis B virus (HBV) DNA in serum is a useful method for the monitoring of HBV replication. We attempted to develop a quantitative assay system for HBV DNA that is more sensitive, accurate, and reproducible than existing systems. We detected HBV DNA by real-time detection PCR (RTD-PCR) based on Taq Man chemistry. The efficacy of this assay was evaluated by quantitatively measuring sequential levels of synthetic DNA and DNA in clinical serum samples. The detection limit of this system was as few as 10 DNA copies/reaction. A linear standard curve was obtained between 10(1) and 10(8) DNA copies/reaction. The coefficient of variation for both intra- and interexperimental variability indicated remarkable reproducibility. This system detected HBV DNA in 100% of chronic hepatitis B patients tested and never detected HBV DNA in healthy volunteers who were negative for HBV markers. These observations suggest that RTD-PCR is an excellent candidate for a standard HBV quantification method.
血清中乙型肝炎病毒(HBV)DNA的定量分析是监测HBV复制的一种有用方法。我们试图开发一种用于HBV DNA的定量检测系统,该系统比现有系统更灵敏、准确且可重复。我们基于Taq Man化学原理通过实时检测PCR(RTD-PCR)检测HBV DNA。通过定量测量合成DNA和临床血清样本中DNA的连续水平来评估该检测方法的有效性。该系统的检测限低至10个DNA拷贝/反应。在10(1)至10(8)个DNA拷贝/反应之间获得了线性标准曲线。实验内和实验间变异的变异系数表明具有显著的可重复性。该系统在100%检测的慢性乙型肝炎患者中检测到了HBV DNA,而在HBV标志物呈阴性的健康志愿者中从未检测到HBV DNA。这些观察结果表明,RTD-PCR是标准HBV定量方法的极佳候选者。