Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2020 Oct:549:1-4.
doi: 10.1016/j.virol.2020.07.006. Epub 2020 Jul 29.

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

Tao Wu  1 , Yiyue Ge  2 , Kangchen Zhao  2 , Xiaojuan Zhu  2 , Yin Chen  2 , Bin Wu  2 , Fengcai Zhu  2 , Baoli Zhu  3 , Lunbiao Cui  2
Affiliations
Comparative Study

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

Tao Wu et al. Virology. 2020 Oct.

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.

Keywords: Nucleocapsid protein; Real-time RT-PCR; Real-time RT-RAA; SARS-CoV-2.

PubMed Disclaimer

Conflict of interest statement

None.

Figures

Fig. 1
Fig. 1
Amplification plots of real-time RT-RAA for 10-fold serial dilutions of plasmid.
Fig. 2
Fig. 2
Specificity of the real-time RT-RAA assay for SARS-Cov-2.
See this image and copyright information in PMC

References

    1. Armstrong G.L., Maccannell D., Taylor J., Carleton H.A., Neuhaus E.B., Bradbury R.S., Posey J.E., Gwinn M. Pathogen genomics in public health. N. Engl. J. Med. 2019;381:2569–2580. - PMC - PubMed
    1. Chen C., Li X., Li G., Zhao L., Duan S., Yan T., Feng Z., Ma X. Use of a rapid reverse-transcription recombinase aided amplification assay for respiratory syncytial virus detection. Diagn. Microbiol. Infect. Dis. 2018;90:90–95. - PubMed
    1. Chen N., Zhou M., Dong X., Qu J., Gong F., Han Y., Qiu Y., Wang J., Liu Y., Wei Y. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study. Lancet. 2020;395:507–513. - PMC - PubMed
    1. Corman V.M., Landt O., Kaiser M., Molenkamp R., Meijer A., Chu D.K., Bleicker T., Brunink S., Schneider J., Schmidt M.L. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25:2000045. - PMC - PubMed
    1. Guo L., Wei D., Zhang X., Wu Y., Li Q., Zhou M., Qu J. Clinical features predicting mortality risk in patients with viral pneumonia: the MuLBSTA score. Front. Microbiol. 2019;10 - PMC - PubMed

Publication types

MeSH terms