Performance evaluation of real-time RT-PCR assays for 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 detection of severe acute respiratory syndrome coronavirus-2 developed by the National Institute of Infectious Diseases, Japan.
复制标题
日本国立传染病研究所开发的严重急性呼吸综合征冠状病毒 2 型实时 RT-PCR 检测方法的性能评估。
DOI：
10.7883/yoken.jjid.2020.1079
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发表时间：
2021
影响因子：
2.2
通讯作者：
M. Takeda
中科院分区：
文献类型：
--
作者：
K. Shirato;Yuriko Tomita;H. Katoh;Souichi Yamada;S. Fukushi;S. Matsuyama;M. Takeda
2019 年 12 月中国武汉爆发 2019 年冠状病毒病后不久，日本国家传染病研究所 (NIID) 制定了实时 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 Charité 的核衣壳 (Sarbeco-N) 和 NIID 的核衣壳 (NIID-N2) 检测。在接下来的几个月中，SARS-CoV-2 的传播导致了全球大流行，各种 SARS-CoV-2 序列被注册到公共数据库，例如共享所有流感数据的全球倡议 (GISAID)。在本研究中，我们评估了新开发的 S2 测定 (NIID-S2) 来取代 Sarbeco-N 测定，以及 NIID-N2 和 NIID-S2 测定的性能，涉及引物/探针目标区域中的错配。我们发现 NIID-S2 组的分析灵敏度和特异性与 NIID-N2 测定相当，并且临床标本的检出率与 NIID-N2 测定相同。此外，在可用序列（约 192,000 个）中，NIID-N2 和 NIID-S2 组分别有 2.6% 和 1.2% 的错配序列，尽管大多数错配并不影响扩增效率，但 NIID-N2 正向引物的 3' 端除外。这些发现表明，在新开发的 NIID-S2 套件的支持下，先前开发的 NIID-N2 检测仍然适用于检测 SARS-CoV-2。
Soon after the December 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 Charité's nucleocapsid (Sarbeco-N) and NIID's nucleocapsid (NIID-N2) assays. During the following months, SARS-CoV-2 spread causing a global pandemic, and a variety of SARS-CoV-2 sequences were registered to public databases, such as the Global Initiative on Sharing All Influenza Data (GISAID). In this study, we evaluated the newly developed S2 assay (NIID-S2) to replace the Sarbeco-N assay and the performance of NIID-N2 and NIID-S2 assays, referring mismatches in the primer/probe targeted region. We found the analytical sensitivity and specificity of the NIID-S2 set were comparable to the NIID-N2 assay, and the detection rate for clinical specimens was identical to that of the NIID-N2 assay. Furthermore, among available sequences (approximately 192,000), the NIID-N2 and NIID-S2 sets had 2.6% and 1.2% mismatched sequences, respectively, although most of these mismatches did not affect the amplification efficiency, with the exception of the 3' end of the NIID-N2 forward primer. These findings indicate that the previously developed NIID-N2 assay remains suitable for the detection SARS-CoV-2 with support of the newly developed NIID-S2 set.
影响因子：
19
作者：
Corman, Victor M.;Landt, Olfert;Drosten, Christian
通讯作者：
Drosten, Christian