Performance Evaluation of Real -Time RT-PCR Assays for the Detection of Severe Acute Respiratory Syndrome Coronavirus-2 Developed by the National Institute of Infectious Diseases, Japan
Performance Evaluation of Real -Time RT-PCR Assays for the Detection of Severe Acute Respiratory Syndrome Coronavirus-2 Developed by the National Institute of Infectious Diseases, Japan
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DOI：
10.7883/yoken.hid.2020.1079
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发表时间：
2021-09-01
影响因子：
2.2
通讯作者：
Takeda, Makoto
中科院分区：
文献类型：
--
作者：
Shirato, Kazuya;Tomita, Yuriko;Takeda, Makoto
2019年中国武汉爆发2019冠状病毒病后不久，日本国立传染病研究所（NIID）建立了一种实时RT-PCR检测严重急性呼吸综合征冠状病毒（SARS-CoV-2）的方案。该方案使用Charite核衣壳（Sarbeco-N）和NIID核衣壳（NIID-N2）测定。在接下来的几个月里，SARS-CoV-2传播并导致全球大流行，各种SARS-CoV-2序列在公共数据库中注册，例如全球共享所有流感数据倡议（GISAID）。在本研究中，我们评价了新开发的S2检测试剂盒（NIID-S2），以取代Sarbeco-N检测试剂盒，以及NIID-N2和NIID-S2检测试剂盒的性能，涉及引物/探针靶向区域的错配。结果表明，NIID-S2试剂盒的灵敏度和特异性与NIID-N2试剂盒相当，对临床标本的检出率与NIID-N2试剂盒相同。此外，在可用序列（约192，000个）中，NIID-N2和NIID-S2组分别具有2.6%和1.2 A错配序列，尽管这些错配中的大多数不影响扩增效率，除了NIID-N2正向引物的3'末端。这些发现表明，先前开发的NIID-N2检测试剂盒适用于检测SARS-CoV-2，并得到新开发的NIID-S2试剂盒的支持。
Soon after the 2019 outbreak of coronavirus disease 2019 in Wuhan, China, a protocol for real-time RT-PCR assay detection of severe acute respiratory syndrome coronavirus (SARS-CoV-2) was established by the National Institute of Infectious Diseases (NIID) in Japan. The protocol used Charite's nucleocapsid (Sarbeco-N) and NIID nucleocapsid (NIID-N2) assays. During the following months, SARS-CoV-2 spread and caused a global pandemic, and various SARS-CoV-2 sequences were registered in public databases, such as the Global Initiative on Sharing All Influenza Data (GISAID). In this study, we evaluated the S2 assay (NIID-S2) that was newly developed to replace the Sarbeco-N assay and the performance of the NIID-N2 and NIID-S2 assays, referring to mismatches in the primer/ probe targeted region. We found that the analytical sensitivity and specificity of the NIID-S2 set were comparable to those of the NIID-N2 assay, and the cetection rate for clinical specimens was identical to that of the NIID-N2 assay. Furthermore, among the available sequences (approximately 192,000), the NIID-N2 and NIID-S2 sets had 2.6% and 1.2 A mismatched sequences, respectively, although most of these mismatches did not affect the amplification efficiency, except the 3' end of the NIID-N2 forward primer. These findings indicate that the previously developed NIID-N2 assay is suitable for the detection of SARS-CoV-2 with support from the newly developed NIID-S2 set.