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; Yiyue Ge; Kangchen Zhao; Xiaojuan Zhu; Yin Chen; Bin Wu; Fengcai Zhu; Baoli Zhu; Lunbiao Cui

doi:10.1016/j.virol.2020.07.006

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 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: [email protected].
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.

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.

Publication types

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

MeSH terms

Betacoronavirus / genetics*
Betacoronavirus / isolation & purification
COVID-19
COVID-19 Testing
COVID-19 Vaccines
China / epidemiology
Clinical Laboratory Techniques / methods*
Clinical Laboratory Techniques / statistics & numerical data
Coronavirus Infections / diagnosis
Coronavirus Infections / epidemiology
Coronavirus Infections / virology*
Coronavirus Nucleocapsid Proteins
Genes, Viral*
Humans
Limit of Detection
Nucleic Acid Amplification Techniques / methods*
Nucleic Acid Amplification Techniques / statistics & numerical data
Nucleocapsid Proteins / genetics*
Pandemics
Phosphoproteins
Pneumonia, Viral / diagnosis
Pneumonia, Viral / epidemiology
Pneumonia, Viral / virology*
RNA, Viral / analysis
RNA, Viral / genetics
Real-Time Polymerase Chain Reaction / methods
Real-Time Polymerase Chain Reaction / statistics & numerical data
Recombinases
Reverse Transcriptase Polymerase Chain Reaction / methods
Reverse Transcriptase Polymerase Chain Reaction / statistics & numerical data
SARS-CoV-2
Sensitivity and Specificity

Substances

COVID-19 Vaccines
Coronavirus Nucleocapsid Proteins
Covid-19 aAPC vaccine
Nucleocapsid Proteins
Phosphoproteins
RNA, Viral
Recombinases
nucleocapsid phosphoprotein, SARS-CoV-2