Rashed-Ul Islam S M, Jahan Munira, Tabassum Shahina
Department of Virology, Bangabandhu Sheikh Mujib Medical University, Shahbag, Dhaka, Bangladesh.
Euroasian J Hepatogastroenterol. 2015 Jan-Jun;5(1):11-15. doi: 10.5005/jp-journals-10018-1121. Epub 2015 Jan 6.
Virological monitoring is the best predictor for the management of chronic hepatitis B virus (HBV) infections. Consequently, it is important to use the most efficient, rapid and cost-effective testing systems for HBV DNA quantification. The present study compared the performance characteristics of a one-step HBV polymerase chain reaction (PCR) the two-step HBV PCR method for quantification of HBV DNA from clinical samples. A total of 100 samples consisting of 85 randomly selected samples from patients with chronic hepatitis B (CHB) and 15 samples from apparently healthy individuals were enrolled in this study. Of the 85 CHB clinical samples tested, HBV DNA was detected from 81% samples by one-step PCR method with median HBV DNA viral load (VL) of 7.50 × 10 lU/ml. In contrast, 72% samples were detected by the two-step PCR system with median HBV DNA of 3.71 × 10 lU/ml. The one-step method showed strong linear correlation with two-step PCR method (r = 0.89; p < 0.0001). Both methods showed good agreement at Bland-Altman plot, with a mean difference of 0.61 log IU/ml and limits of agreement of -1.82 to 3.03 log IU/ml. The intra-assay and interassay coefficients of variation (CV%) of plasma samples (4-7 log IU/ml) for the one-step PCR method ranged between 0.33 to 0.59 and 0.28 to 0.48 respectively, thus demonstrating a high level of concordance between the two methods. Moreover, elimination of the DNA extraction step in the one-step PCR kit allowed time-efficient and significant labor and cost savings for the quantification of HBV DNA in a resource limited setting.
Rashed-Ul Islam SM, Jahan M, Tabassum S. Evaluation of a Rapid One-step Real-time PCR Method as a High-throughput Screening for Quantification of Hepatitis B Virus DNA in a Resource-limited Setting. Euroasian J Hepato-Gastroenterol 2015;5(1):11-15.
病毒学监测是慢性乙型肝炎病毒（HBV）感染管理的最佳预测指标。因此，使用最有效、快速且具有成本效益的检测系统进行HBV DNA定量非常重要。本研究比较了一步法HBV聚合酶链反应（PCR）和两步法HBV PCR对临床样本中HBV DNA定量的性能特征。本研究共纳入了100份样本，其中包括从慢性乙型肝炎（CHB）患者中随机选取的85份样本以及15份来自表面健康个体的样本。在检测的85份CHB临床样本中，一步法PCR检测出81%的样本存在HBV DNA，HBV DNA病毒载量（VL）中位数为7.50×10 IU/ml。相比之下，两步法PCR系统检测出72%的样本，HBV DNA中位数为3.71×10 IU/ml。一步法与两步法PCR方法显示出强线性相关性（r = 0.89；p < 0.0001）。两种方法在布兰德 - 奥特曼图上显示出良好的一致性，平均差异为0.61 log IU/ml，一致性界限为 -1.82至3.03 log IU/ml。一步法PCR对血浆样本（4 - 7 log IU/ml）的批内和批间变异系数（CV%）分别在0.33至0.59和0.28至0.48之间，从而证明两种方法具有高度一致性。此外，一步法PCR试剂盒省去了DNA提取步骤，在资源有限的情况下，可高效省时，并显著节省人力和成本用于HBV DNA定量。
Rashed-Ul Islam SM，Jahan M，Tabassum S。在资源有限环境中评估一种快速一步实时PCR方法作为高通量筛选定量乙型肝炎病毒DNA的方法。《欧亚肝脏胃肠病学杂志》2015；5(1):11 - 15。
