Reverse Transcription Recombinase-Aided Amplification Assay With Lateral Flow Dipstick Assay for Rapid Detection of 2019 Novel Coronavirus

作者：Yuzhong Zheng, Jiang‐Tao Chen, Jian Li, Xianjing Wu, Jin-Zhou Wen, Xiangzhi Liu, Li‐Yun Lin, Xue‐Yan Liang, Huiying Huang, Guangcai Zha, Peikui Yang, Liejun Li, Tianyu Zhong, Long Liu, Weijia Cheng, Xiaonan Song, Min Lin · 发表于：Frontiers in Cellular and Infection Microbiology · 年份：2021 · DOI：10.3389/fcimb.2021.613304 · 被引用次数：62 · 研究领域：SARS-CoV-2 detection and testing、Biosensors and Analytical Detection、Molecular Biology Techniques and Applications

Background The emerging Coronavirus Disease-2019 (COVID-19) has challenged the public health globally. With the increasing requirement of detection for SARS-CoV-2 outside of the laboratory setting, a rapid and precise Point of Care Test (POCT) is urgently needed. Methods Targeting the nucleocapsid (N) gene of SARS-CoV-2, specific primers, and probes for reverse transcription recombinase-aided amplification coupled with lateral flow dipstick (RT-RAA/LFD) platform were designed. For specificity evaluation, it was tested with human coronaviruses, human influenza A virus, influenza B viruses, respiratory syncytial virus, and hepatitis B virus, respectively. For sensitivity assay, it was estimated by templates of recombinant plasmid and pseudovirus of SARS-CoV-2 RNA. For clinical assessment, 100 clinical samples (13 positive and 87 negatives for SARS-CoV-2) were tested via quantitative reverse transcription PCR (RT-qPCR) and RT-RAA/LFD, respectively. Results The limit of detection was 1 copies/μl in RT-RAA/LFD assay, which could be conducted within 30 min at 39°C, without any cross-reaction with other human coronaviruses and clinical respiratory pathogens. Compared with RT-qPCR, the established POCT assay offered 100% specificity and 100% sensitivity in the detection of clinical samples. Conclusion This work provides a convenient POCT tool for rapid screening, diagnosis, and monitoring of suspected patients in SARS-CoV-2 endemic areas.