Faculty of Biological Sciences, School of Molecular and Cellular Biology, University of Leeds, Leeds, UK.
Faculty of Clinical Sciences, Department of Epidemiology and Community Health, College of Health Sciences, University of Ilorin, Ilorin, Nigeria.
J Med Virol. 2017 Nov;89(11):1944-1951. doi: 10.1002/jmv.24877. Epub 2017 Jul 17.
The Hepatitis B surface antigen (HBsAg) is the hallmark of HBV infection. Detection of antibodies to HBs and the core (ie, HBsAg and HBcAb) are primary serological algorithms in the laboratory diagnosis of HBV. Detection of HBsAg DNA is an important supplement to serological diagnosis especially in clinical cases. Simultaneous amplification of internal cellular controls is a good indicator of sample quality. Human β-globin is a well characterized housekeeping gene (HKG) that is often applied as internal controls (IC) in molecular diagnosis. In this study, individual plasmid clones of the human β-globin and HBs genes were constructed. These plasmid constructs have been applied to characterize a multiplex PCR assays for HBs and β-globin genes. The findings suggest detection limits of less than 10 genome copies of either template In vitro using conventional and multiplex PCR conditions. Under the multiplex conditions, co-amplification of β-globin and HBsAg DNA had a resultant effect on assay sensitivity. This study further highlights the importance of molecular diagnosis in HBV infectious individuals. If fully optimized, this assay could provide a possible diagnostic complement to serological detection in developing countries.
乙型肝炎表面抗原 (HBsAg) 是乙型肝炎病毒 (HBV) 感染的标志。检测乙型肝炎表面抗体 (HBs) 和核心抗体（即 HBsAg 和 HBcAb）是实验室诊断乙型肝炎的主要血清学算法。检测 HBsAg DNA 是血清学诊断的重要补充，特别是在临床病例中。同时扩增内参细胞控制物是样本质量的良好指标。人β-球蛋白是一种特征明确的管家基因 (HKG)，常用于分子诊断中的内参 (IC)。在本研究中，构建了人β-球蛋白和 HBs 基因的单个质粒克隆。这些质粒构建物已应用于描述用于 HBs 和 β-球蛋白基因的多重 PCR 分析。研究结果表明，在常规和多重 PCR 条件下，体外检测到的模板的检测限低于 10 个基因组拷贝。在多重条件下，β-球蛋白和 HBsAg DNA 的共扩增对检测敏感性有影响。本研究进一步强调了分子诊断在乙型肝炎病毒感染者中的重要性。如果经过充分优化，该检测方法可能为发展中国家的血清学检测提供一种可能的诊断补充。
