1. Reverse Transcription Recombinase-Aided Amplification Assay With Lateral Flow Dipstick Assay for Rapid Detection of 2019 Novel Coronavirus.
- Author
-
Zheng, Yu-Zhong, Chen, Jiang-Tao, Li, Jian, Wu, Xian-Jing, Wen, Jin-Zhou, Liu, Xiang-Zhi, Lin, Li-Yun, Liang, Xue-Yan, Huang, Hui-Ying, Zha, Guang-Cai, Yang, Pei-Kui, Li, Lie-Jun, Zhong, Tian-Yu, Liu, Long, Cheng, Wei-Jia, Song, Xiao-Nan, and Lin, Min
- Subjects
SARS-CoV-2 ,CORONAVIRUSES ,RESPIRATORY syncytial virus ,COVID-19 ,INFLUENZA viruses ,HEPATITIS B virus - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF