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

doi:10.1016/j.virol.2020.07.006

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 Wu¹, Yiyue Ge², Kangchen Zhao², Xiaojuan Zhu², Yin Chen², Bin Wu², Fengcai Zhu², Baoli Zhu³, Lunbiao Cui²

Affiliations

¹ From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China. Electronic address: lbcui@jscdc.cn.
² From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.
³ From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China; From Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, 211166, China.

PMID: 32758712
PMCID: PMC7388781
DOI: 10.1016/j.virol.2020.07.006

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.

. 2020 Oct:549:1-4.

doi: 10.1016/j.virol.2020.07.006. Epub 2020 Jul 29.

Authors

Tao Wu¹, Yiyue Ge², Kangchen Zhao², Xiaojuan Zhu², Yin Chen², Bin Wu², Fengcai Zhu², Baoli Zhu³, Lunbiao Cui²

Affiliations

¹ From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China. Electronic address: lbcui@jscdc.cn.
² From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.
³ From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China; From Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, 211166, China.

PMID: 32758712
PMCID: PMC7388781
DOI: 10.1016/j.virol.2020.07.006

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**
Amplification plots of real-time RT-RAA for 10-fold serial dilutions of plasmid.

**Fig. 2**
Specificity of the real-time RT-RAA assay for SARS-Cov-2.

See this image and copyright information in PMC

Cited by

The Role of Imaging in the Detection of Non-COVID-19 Pathologies during the Massive Screening of the First Pandemic Wave.
Canivet P, Desir C, Thys M, Henket M, Frix AN, Ernst B, Walsh S, Occhipinti M, Vos W, Maes N, Canivet JL, Louis R, Meunier P, Guiot J. Canivet P, et al. Diagnostics (Basel). 2022 Jun 28;12(7):1567. doi: 10.3390/diagnostics12071567. Diagnostics (Basel). 2022. PMID: 35885473 Free PMC article.
A Reverse Transcription Recombinase-Aided Amplification Method for Rapid and Point-of-Care Detection of SARS-CoV-2, including Variants.
Li F, He P, Xiong D, Lou Y, Pu Q, Zhang H, Zhang H, Yu J. Li F, et al. Viruses. 2021 Sep 19;13(9):1875. doi: 10.3390/v13091875. Viruses. 2021. PMID: 34578456 Free PMC article.
Suitcase Lab for Rapid Detection of SARS-CoV-2 Based on Recombinase Polymerase Amplification Assay.
El Wahed AA, Patel P, Maier M, Pietsch C, Rüster D, Böhlken-Fascher S, Kissenkötter J, Behrmann O, Frimpong M, Diagne MM, Faye M, Dia N, Shalaby MA, Amer H, Elgamal M, Zaki A, Ismail G, Kaiser M, Corman VM, Niedrig M, Landt O, Faye O, Sall AA, Hufert FT, Truyen U, Liebert UG, Weidmann M. El Wahed AA, et al. Anal Chem. 2021 Feb 2;93(4):2627-2634. doi: 10.1021/acs.analchem.0c04779. Epub 2021 Jan 20. Anal Chem. 2021. PMID: 33471510 Free PMC article.
Sensitive methods for detection of SARS-CoV-2 RNA.
Chen X, Xia S. Chen X, et al. Methods Microbiol. 2022;50:1-26. doi: 10.1016/bs.mim.2021.06.001. Epub 2021 Jul 16. Methods Microbiol. 2022. PMID: 38620918 Free PMC article.
Epidemiological investigation of swine Japanese encephalitis virus based on RT-RAA detection method.
Nie M, Zhou Y, Li F, Deng H, Zhao M, Huang Y, Jiang C, Sun X, Xu Z, Zhu L. Nie M, et al. Sci Rep. 2022 Jun 7;12(1):9392. doi: 10.1038/s41598-022-13604-4. Sci Rep. 2022. PMID: 35672440 Free PMC article.

See all "Cited by" articles

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

Actions
Actions
Actions

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

LinkOut - more resources

Full Text Sources
Medical
- MedlinePlus Health Information
Miscellaneous
- NCI CPTAC Assay Portal

[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

[2] 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

[3] 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

[4] 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

[5] 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

[6] 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

[7] 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

[8] 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

[9] 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

[10] 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

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

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

Affiliations

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

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

Publication types

MeSH terms

Substances

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

Publication types

MeSH terms

Substances

Related information

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous