Department of Bacteriology, Capital Institute of Pediatrics, Beijing 100020, P. R. China.
Treatment and Research Center for Infectious Diseases, the Fifth Medical Center of PLA General Hospital, Beijing 100039, P. R. China.
Anal Chem. 2020 Jul 21;92(14):9699-9705. doi: 10.1021/acs.analchem.0c01032. Epub 2020 Jul 10.
A novel coronavirus (SARS-CoV-2) was recently identified in patients with acute respiratory disease and spread quickly worldwide. A specific and rapid diagnostic method is important for early identification. The reverse-transcription recombinase-aided amplification (RT-RAA) assay is a rapid detection method for several pathogens. Assays were performed within 5-15 min as a one-step single tube reaction at 39 °C. In this study, we established two RT-RAA assays for the and gene of SARS-CoV-2 using clinical specimens for validation. The analytical sensitivity of the RT-RAA assay was 10 copies for the and one copy for the gene per reaction. Cross-reactions were not observed with any of the other respiratory pathogens. A 100% agreement between the RT-RAA and real-time PCR assays was accomplished after testing 120 respiratory specimens. These results demonstrate that the proposed RT-RAA assay will be beneficial as it is a faster, more sensitive, and more specific tool for the detection of SARS-CoV-2.
一种新型冠状病毒（SARS-CoV-2）最近在急性呼吸道疾病患者中被发现，并迅速在全球范围内传播。一种特异性和快速的诊断方法对于早期识别非常重要。逆转录重组酶辅助扩增（RT-RAA）检测是针对几种病原体的快速检测方法。该检测在 39°C 下进行一步式单管反应，可在 5-15 分钟内完成。在这项研究中，我们使用临床标本建立了两种用于 SARS-CoV-2 的 和 基因的 RT-RAA 检测方法进行验证。该 RT-RAA 检测的分析灵敏度为每个反应 10 拷贝的 和 1 拷贝的 基因。与任何其他呼吸道病原体均无交叉反应。对 120 份呼吸道标本进行检测后，RT-RAA 与实时 PCR 检测方法的一致性达到 100%。这些结果表明，拟议的 RT-RAA 检测方法将是有益的，因为它是一种更快、更敏感、更特异的检测 SARS-CoV-2 的工具。
