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A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).
Virology. 2020 10; 549:1-4.V

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.

Authors+Show Affiliations

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.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.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.From Institute of Pathogenic Microbiology, NHC Key Laboratories of Enteric Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32758712

Citation

Wu, Tao, et al. "A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of N Gene of Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2)." Virology, vol. 549, 2020, pp. 1-4.
Wu T, Ge Y, Zhao K, et al. A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Virology. 2020;549:1-4.
Wu, T., Ge, Y., Zhao, K., Zhu, X., Chen, Y., Wu, B., Zhu, F., Zhu, B., & Cui, L. (2020). A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Virology, 549, 1-4. https://doi.org/10.1016/j.virol.2020.07.006
Wu T, et al. A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of N Gene of Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2). Virology. 2020;549:1-4. PubMed PMID: 32758712.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). AU - Wu,Tao, AU - Ge,Yiyue, AU - Zhao,Kangchen, AU - Zhu,Xiaojuan, AU - Chen,Yin, AU - Wu,Bin, AU - Zhu,Fengcai, AU - Zhu,Baoli, AU - Cui,Lunbiao, Y1 - 2020/07/29/ PY - 2020/04/26/received PY - 2020/07/09/revised PY - 2020/07/09/accepted PY - 2020/8/8/pubmed PY - 2020/9/25/medline PY - 2020/8/8/entrez KW - Nucleocapsid protein KW - Real-time RT-PCR KW - Real-time RT-RAA KW - SARS-CoV-2 SP - 1 EP - 4 JF - Virology JO - Virology VL - 549 N2 - 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. SN - 1096-0341 UR - https://www.unboundmedicine.com/medline/citation/32758712/A_reverse_transcription_recombinase_aided_amplification_assay_for_the_rapid_detection_of_N_gene_of_severe_acute_respiratory_syndrome_coronavirus_2_SARS_CoV_2__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0042-6822(20)30133-1 DB - PRIME DP - Unbound Medicine ER -