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.

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