Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples

Sci Rep. 2021 Jan 19;11(1):1820. doi: 10.1038/s41598-020-80352-8.

Abstract

RT-LAMP detection of SARS-CoV-2 has been shown to be a valuable approach to scale up COVID-19 diagnostics and thus contribute to limiting the spread of the disease. Here we present the optimization of highly cost-effective in-house produced enzymes, and we benchmark their performance against commercial alternatives. We explore the compatibility between multiple DNA polymerases with high strand-displacement activity and thermostable reverse transcriptases required for RT-LAMP. We optimize reaction conditions and demonstrate their applicability using both synthetic RNA and clinical patient samples. Finally, we validate the optimized RT-LAMP assay for the detection of SARS-CoV-2 in unextracted heat-inactivated nasopharyngeal samples from 184 patients. We anticipate that optimized and affordable reagents for RT-LAMP will facilitate the expansion of SARS-CoV-2 testing globally, especially in sites and settings where the need for large scale testing cannot be met by commercial alternatives.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • Hot Temperature
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / metabolism
  • RNA-Directed DNA Polymerase / metabolism
  • Reagent Kits, Diagnostic
  • SARS-CoV-2 / genetics*
  • SARS-CoV-2 / isolation & purification
  • Sensitivity and Specificity
  • Virus Inactivation

Substances

  • RNA, Viral
  • Reagent Kits, Diagnostic
  • RNA-Directed DNA Polymerase

Supplementary concepts

  • LAMP assay