Assessment of Real-Time RT-PCR Kits for SARS-CoV-2 Detection
Assessment of Real-Time RT-PCR Kits for SARS-CoV-2 Detection
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DOI：
10.7883/yoken.jjid.2020.108
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发表时间：
2020-09-01
影响因子：
2.2
通讯作者：
Takeda, Makoto
中科院分区：
文献类型：
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作者：
Okamaoto, Kiyoko;Shirato, Kazuya;Takeda, Makoto
2019年12月在武汉（中国）爆发的由严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）疫情目前正在全球范围内迅速蔓延。我们最近报道了一种基于实时逆转录酶PCR（RT-PCR）检测的SARS-CoV-2诊断实验室方案，使用两组引物，N和N2。2020年1月30日至31日，这些检测的方案和试剂被分发到日本全国各地的地方公共卫生机构和检疫站，此后开始了SARS-CoV-2诊断检测。为了进一步验证，使用从COVID-19疑似个体获得的SARS-CoV-2病毒RNA和临床标本，将该检测方法与市售试剂盒进行了比较。LightMix Modular SARS and Wuhan CoV E-gene（LM S&W-E）检测试剂盒对SARS-CoV-2高度敏感，N2组也是如此，因为两种检测试剂盒对临床标本的结果一致。LM S&W-E组靶向SARS-CoV和SARS-CoV-2中E基因的高度保守区域，而N2组被设计为特异性靶向SARS-CoV-2 N基因中的独特区域。因此，N2组对SARS-CoV-2检测表现出高特异性和灵敏度。这些结果表明，使用N和N2组的方案与市售试剂盒相当，因此对于COVID-19的实验室诊断是可靠的。
The coronavirus induced disease 2019 (COVID-19) outbreak caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Wuhan (China) in December 2019 is currently spreading rapidly worldwide. We recently reported a laboratory protocol for the diagnosis of SARS-CoV-2 based on real-time reverse transcriptase PCR (RT-PCR) assays using two primer sets, N and N2. On January 30-31, 2020, the protocol and the reagents for these assays were distributed to the local public health institutes and quarantine depots in Japan nationwide, and thereafter SARS-CoV-2 diagnostic testing was initiated. For further validation, the assays were compared with the commercially available kits using the SARS-CoV-2 viral RNA and clinical specimens obtained from COVID-19-suspected individuals. The LightMix Modular SARS and Wuhan CoV E-gene (LM S&W-E) assay was highly sensitive for the SARS-CoV-2, as was the N2 set, as both the assays showed consistent results for the clinical specimens. While the LM S&W-E set targets the highly conserved region of E gene in the SARS-CoV and SARS-CoV-2, the N2 set was designed to target specifically the unique region in the SARS-CoV-2 N gene. Therefore, the N2 set exhibits high specificity and sensitivity for SARS-CoV-2 detection. These results indicate that the protocol using the N and N2 sets is comparable to the commercially available kits, and thus is reliable for laboratory diagnosis of COVID-19.