Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2

BMC Infect Dis. 2021 Nov 17;21(1):1162. doi: 10.1186/s12879-021-06876-0.

Abstract

Background: Current assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from nasopharyngeal swab samples that bypasses the RNA purification step.

Methods: In the current project, we have described two extraction-free reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of SARS-CoV-2 by using E gene and RdRp gene as the targets.

Results: Here, results showed that reverse transcription loop-mediated isothermal amplification assays with 88.4% sensitive (95% CI: 74.9-96.1%) and 67.4% sensitive (95% CI: 51.5-80.9%) for E gene and RdRp gene, respectively.

Conclusion: Without the need of RNA purification, our developed RT-LAMP assays for direct detection of SARS-CoV-2 from nasopharyngeal swab samples could be turned into alternatives to qRT-PCR for rapid screening.

Keywords: Coronaviruses; Isothermal detection; RT-LAMP; Rapid diagnosis; SARS-CoV-2.

MeSH terms

  • COVID-19*
  • Humans
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • RNA, Viral / genetics
  • Reverse Transcription
  • SARS-CoV-2*
  • Sensitivity and Specificity

Substances

  • RNA, Viral

Supplementary concepts

  • LAMP assay