RT-qPCR Testing of SARS-CoV-2: A Primer

Int J Mol Sci. 2020 Apr 24;21(8):3004. doi: 10.3390/ijms21083004.

Abstract

Testing for the presence of coronavirus is an essential diagnostic tool for monitoring and managing the current COVID-19 pandemic. The only reliable test in current use for testing acute infection targets the genome of SARS-CoV-2, and the most widely used method is quantitative fluorescence-based reverse transcription polymerase chain reaction (RT-qPCR). Despite its ubiquity, there is a significant amount of uncertainty about how this test works, potential throughput and reliability. This has resulted in widespread misrepresentation of the problems faced using this test during the current COVID-19 epidemic. This primer provides simple, straightforward and impartial information about RT-qPCR.

Keywords: COVID-19; SARS; pandemic; real-time fluorescence PCR; reverse transcription.

MeSH terms

  • Betacoronavirus / genetics
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • COVID-19 Testing
  • Clinical Laboratory Techniques / methods*
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / virology
  • DNA Primers*
  • Pandemics
  • Pneumonia, Viral / diagnosis
  • Pneumonia, Viral / virology
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Real-Time Polymerase Chain Reaction / methods*
  • SARS-CoV-2

Substances

  • DNA Primers
  • RNA, Viral