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一种逆转录重组酶辅助扩增检测法,用于快速检测严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的 N 基因。

A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).

机构信息

From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.

From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.

出版信息

Virology. 2020 Oct;549:1-4. doi: 10.1016/j.virol.2020.07.006. Epub 2020 Jul 29.

Abstract

The current outbreak of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was reported in China firstly. A rapid, highly sensitive, specific, and simple operational method was needed for the detection of SARS-CoV-2. Here, we established a real-time reverse-transcription recombinase-aided amplification assay (RT-RAA) to detect SARS-CoV-2 rapidly. The primers and probe were designed based on the nucleocapsid protein gene (N gene) sequence of SARS-CoV-2. The detection limit was 10 copies per reaction in this assay, which could be conducted within 15 min at a constant temperature (39 °C), without any cross-reactions with other respiratory tract pathogens, such as other coronaviruses. Furthermore, compared with commercial real-time RT-PCR assay, it showed a kappa value of 0.959 (p < 0.001) from 150 clinical specimens. These results indicated that this real-time RT-RAA assay may be a valuable tool for detecting SARS-CoV-2.

摘要

目前由严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)引起的 2019 年冠状病毒病(COVID-19)疫情首先在中国报告。需要一种快速、高灵敏度、特异性和简单的操作方法来检测 SARS-CoV-2。在这里,我们建立了一种实时逆转录重组酶辅助扩增检测(RT-RAA)方法来快速检测 SARS-CoV-2。该检测方法基于 SARS-CoV-2 的核衣壳蛋白基因(N 基因)序列设计了引物和探针。该检测方法的检测限为每个反应 10 个拷贝,可在 39°C 恒温下 15 分钟内完成,与其他呼吸道病原体(如其他冠状病毒)无交叉反应。此外,与商业实时 RT-PCR 检测方法相比,从 150 份临床标本中得到的kappa 值为 0.959(p<0.001)。这些结果表明,该实时 RT-RAA 检测方法可能是一种检测 SARS-CoV-2 的有价值的工具。

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