Development of Reverse Transcription Loop-Mediated Isothermal Amplification Assays Targeting Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

J Mol Diagn. 2020 Jun;22(6):729-735. doi: 10.1016/j.jmoldx.2020.03.006. Epub 2020 Apr 7.

Abstract

The coronavirus disease 2019 (COVID-19) pandemic now has >2,000,000 confirmed cases worldwide. COVID-19 is currently diagnosed using quantitative RT-PCR methods, but the capacity of quantitative RT-PCR methods is limited by their requirement of high-level facilities and instruments. We developed and evaluated reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays to detect genomic RNA of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative virus of COVID-19. RT-LAMP assays reported in this study can detect as low as 100 copies of SARS-CoV-2 RNA. Cross-reactivity of RT-LAMP assays to other human coronaviruses was not observed. A colorimetric detection method was adapted for this RT-LAMP assay to enable higher throughput.

MeSH terms

  • Betacoronavirus / genetics*
  • Colorimetry / methods
  • DNA Primers
  • Gentian Violet
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / genetics
  • Reverse Transcription
  • SARS-CoV-2

Substances

  • DNA Primers
  • RNA, Viral
  • Gentian Violet