Liu Chao, Chang Le, Wang Lunan
1National Center for Clinical Laboratories, Beijing Hospital, National Center of Gerontology, No. 1 Dahua Road, Dongdan, Beijing, 100730 People's Republic of China.
2Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, Beijing, People's Republic of China.
Indian J Hematol Blood Transfus. 2019 Jul;35(3):544-550. doi: 10.1007/s12288-018-01064-8. Epub 2019 Jan 18.
To analyze the risk and reason of false-negative HBV DNA results of NAT reagents among blood donations of China and discuss the necessity of two amplification targets for HBV DNA tests among donations. In this study, samples that showed discordant results on two commercially available assay platforms were further detected by established in-house methods based on conserved regions of the HBV genome. The HBV concentration of these samples was determined using two commercially available reagents. The samples with high titers of HBV were detected by an in-house method. The samples showing high Ct differences between two regions in the in-house method were further sequenced and aligned with primers and probes. The results showed that the established method has a good detection performance. The mismatch between reverse primers and sample sequences led to decreased detection capacity of S and C regions by the in-house method, but it could be compensated by another region. Among the false-negative samples detected by commercial reagents, most were because of low titers; however, there were 7 samples with HBV DNA concentrations much higher than the LOD of the commercial reagents, which may be due to mismatch of the primer or probe. This study highlights the potential risk of HBV false-negative detection by commercial NAT reagents. The dual-target assay may be helpful for HBV screening and reduce the risk of false-negative detection.
分析我国献血人群中核酸扩增技术（NAT）试剂检测乙型肝炎病毒（HBV）DNA出现假阴性结果的风险及原因，并探讨献血者HBV DNA检测采用双扩增靶点的必要性。在本研究中，对在两种商用检测平台上结果不一致的样本，采用基于HBV基因组保守区域建立的内部方法进行进一步检测。使用两种商用试剂测定这些样本的HBV浓度。对高滴度HBV样本采用内部方法进行检测。对内部方法中两个区域之间Ct差异较大的样本进一步测序，并与引物和探针进行比对。结果表明，所建立的方法具有良好的检测性能。反向引物与样本序列不匹配导致内部方法对S区和C区的检测能力下降，但可通过另一个区域进行补偿。在商用试剂检测出的假阴性样本中，大多数是由于滴度较低；然而，有7个样本的HBV DNA浓度远高于商用试剂的检测下限，这可能是由于引物或探针不匹配所致。本研究突出了商用NAT试剂检测HBV出现假阴性的潜在风险。双靶点检测可能有助于HBV筛查并降低假阴性检测风险。
