High-quality RNA improves sensitivity of SARS-CoV-2 detection by colorimetric RT-LAMP

Exp Biol Med (Maywood). 2022 Feb;247(3):276-281. doi: 10.1177/15353702211054768. Epub 2021 Dec 14.

Abstract

The global SARS-CoV-2 pandemic requires a rapid, reliable, and user-friendly diagnostic test to help control the spread of the virus. Reverse transcription and quantitative PCR (RT-qPCR) is currently the gold standard method for SARS-CoV-2 detection. Here, we develop a protocol based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and demonstrate increased sensitivity of this technique using fresh RNA extracts compared to RNA samples subjected to freezing/thawing cycles. We further compare RT-LAMP to RT-qPCR and demonstrate that the RT-LAMP approach has high sensitivity in fresh RNA extracts and can detect positive samples with Ct values between 8 and 35.

Keywords: RT-LAMP; SARS-CoV-2; high-quality RNA.

MeSH terms

  • COVID-19 Nucleic Acid Testing / methods*
  • Colorimetry / methods
  • DNA Primers
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / isolation & purification*
  • Reverse Transcription
  • SARS-CoV-2 / genetics
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • RNA, Viral

Supplementary concepts

  • LAMP assay