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 , Lin Min 

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

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 11 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2021.613304

DOI=10.3389/fcimb.2021.613304

ISSN=2235-2988

ABSTRACT=Background
The emerging Coronavirus Disease-2019 (COVID-19) has challenged the global public health community. With increasing requirement of detection for SARS-CoV-2 outside of the laboratory setting, a rapid and accurate Point of Care Test (POCT) is required.
Methods
Targeting the nucleocapsid (N) gene of SARS-CoV-2, specific primers and probe for reverse transcription recombinase-aided amplification coupled with lateral flow dipstick (RT-RAA/LFD) platform were designed. Specificity of the RT-RAA/LFD assays was tested with human coronaviruses, human influenza A virus, influenza B viruses, respiratory syncytial virus and hepatitis B virus. Sensitivity of the RT-RAA/LFD assay was evaluated by using recombinant plasmid and SARS-CoV-2 RNA transcribed in vitro reference material. Finally, 100 clinical samples (13 positive and 87 negative for SARS-CoV-2) were tested using both quantitative reverse transcription PCR (RT-qPCR) and RT-RAA/LFD.
Results
The detection limit was 1 copies/μL in RT-RAA/LFD assay, which could be conducted within 30 min at 39 °C, without any cross-reactions with other human coronaviruses and clinical respiratory pathogens. Compared with RT-qPCR, the RT-RAA/LFD assay showed 100% specificity and 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.