Portilho Moyra Machado, Baptista Marcia Leite, da Silva Messias, de Sousa Paulo Sérgio Fonseca, Lewis-Ximenez Lia Laura, Lampe Elisabeth, Villar Livia Melo
Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
Laboratory of Enteroviruses, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
J Virol Methods. 2015 Oct;223:40-4. doi: 10.1016/j.jviromet.2015.07.010. Epub 2015 Jul 26.
The aim of the present study was to evaluate the performance of three in-house PCR techniques for HBV DNA detection and compare it with commercial quantitative methods to evaluate the usefulness of in-house methods for HBV diagnosis. Three panels of HBsAg reactive sera samples were evaluated: (i) 50 samples were examined using three methods for in-house qualitative PCR and the Cobas Amplicor HBV Monitor Assay; (ii) 87 samples were assayed using in-house semi-nested PCR and the Cobas TaqMan HBV test; (iii) 11 serial samples obtained from 2 HBV-infected individuals were assayed using the Cobas Amplicor HBV test and semi-nested PCR. In panel I, HBV DNA was detected in 44 samples using the Cobas Amplicor HBV test, 42 samples using semi-nested PCR (90% concordance with Cobas Amplicor), 22 samples using PCR for the core gene (63.6% concordance) and 29 samples using single-round PCR for the pre-S/S gene (75% concordance). In panel II, HBV DNA was quantified in 78 of the 87 HBsAg reactive samples using Cobas TaqMan but 52 samples using semi-nested PCR (67.8% concordance). HBV DNA was detected in serial samples until the 17th and 26th week after first donation using in-house semi-nested PCR and the Cobas Amplicor HBV test, respectively. In-house semi-nested PCR presented adequate concordance with commercial methods as an alternative method for HBV molecular diagnosis in low-resource settings.
本研究的目的是评估三种内部聚合酶链反应（PCR）技术检测乙型肝炎病毒（HBV）DNA的性能，并将其与商业定量方法进行比较，以评估内部方法在HBV诊断中的实用性。对三组乙型肝炎表面抗原（HBsAg）反应性血清样本进行了评估：（i）使用三种内部定性PCR方法和Cobas Amplicor HBV监测检测法对50份样本进行检测；（ii）使用内部半巢式PCR和Cobas TaqMan HBV检测法对87份样本进行检测；（iii）使用Cobas Amplicor HBV检测法和半巢式PCR对从2名HBV感染个体获得的11份系列样本进行检测。在第一组中，使用Cobas Amplicor HBV检测法在44份样本中检测到HBV DNA，使用半巢式PCR在42份样本中检测到（与Cobas Amplicor的一致性为90%），使用核心基因PCR在22份样本中检测到（一致性为63.6%），使用前S/S基因单轮PCR在29份样本中检测到（一致性为75%）。在第二组中，使用Cobas TaqMan在87份HBsAg反应性样本中的78份中对HBV DNA进行了定量，但使用半巢式PCR在52份样本中进行了定量（一致性为67.8%）。分别使用内部半巢式PCR和Cobas Amplicor HBV检测法在首次献血后第17周和第26周在系列样本中检测到HBV DNA。作为资源匮乏地区HBV分子诊断的替代方法，内部半巢式PCR与商业方法具有足够的一致性。
