Reverse-Transcription Recombinase-Aided Amplification Assay for Rapid Detection of the 2019 Novel Coronavirus (SARS-CoV-2)

Anal Chem. 2020 Jul 21;92(14):9699-9705. doi: 10.1021/acs.analchem.0c01032. Epub 2020 Jul 10.

Abstract

A novel coronavirus (SARS-CoV-2) was recently identified in patients with acute respiratory disease and spread quickly worldwide. A specific and rapid diagnostic method is important for early identification. The reverse-transcription recombinase-aided amplification (RT-RAA) assay is a rapid detection method for several pathogens. Assays were performed within 5-15 min as a one-step single tube reaction at 39 °C. In this study, we established two RT-RAA assays for the S and orf1ab gene of SARS-CoV-2 using clinical specimens for validation. The analytical sensitivity of the RT-RAA assay was 10 copies for the S and one copy for the orf1ab gene per reaction. Cross-reactions were not observed with any of the other respiratory pathogens. A 100% agreement between the RT-RAA and real-time PCR assays was accomplished after testing 120 respiratory specimens. These results demonstrate that the proposed RT-RAA assay will be beneficial as it is a faster, more sensitive, and more specific tool for the detection of SARS-CoV-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / chemistry
  • Bacteria / genetics
  • Betacoronavirus / chemistry*
  • Betacoronavirus / genetics*
  • COVID-19
  • Coronavirus Infections / diagnosis*
  • Cross Reactions
  • DNA Probes
  • Genes, Viral
  • Humans
  • Nucleic Acid Amplification Techniques / methods*
  • Pandemics
  • Plasmids
  • Pneumonia, Viral / diagnosis*
  • Polymerase Chain Reaction / methods*
  • Polyproteins
  • Respiratory Tract Infections / microbiology
  • Respiratory Tract Infections / virology
  • SARS-CoV-2
  • Sensitivity and Specificity
  • Viral Proteins / genetics
  • Viruses / chemistry
  • Viruses / genetics

Substances

  • DNA Probes
  • ORF1ab polyprotein, SARS-CoV-2
  • Polyproteins
  • Viral Proteins