Fast Detection of SARS-CoV-2 RNA Directly from Respiratory Samples Using a Loop-Mediated Isothermal Amplification (LAMP) Test

Viruses. 2021 Apr 29;13(5):801. doi: 10.3390/v13050801.

Abstract

The availability of simple SARS-CoV-2 detection methods is crucial to contain the COVID-19 pandemic. This study examined whether a commercial LAMP assay can reliably detect SARS-CoV-2 genomes directly in respiratory samples without having to extract nucleic acids (NA) beforehand. Nasopharyngeal swabs (NPS, n = 220) were tested by real-time reverse transcription (RT)-PCR and with the LAMP assay. For RT-PCR, NA were investigated. For LAMP, NA from 26 NPS in viral transport medium (VTM) were tested. The other 194 NPS were analyzed directly without prior NA extraction (140 samples in VTM; 54 dry swab samples stirred in phosphate buffered saline). Ten NPS were tested directly by LAMP using a sous-vide cooking unit. The isothermal assay demonstrated excellent specificity (100%) but moderate sensitivity (68.8%), with a positive predictive value of 1 and a negative predictive value of 0.65 for direct testing of NPS in VTM. The use of dry swabs, even without NA extraction, improved the analytical sensitivity; up to 6% of samples showed signs of inhibition. LAMP could be performed successfully with a sous-vide cooking unit. This technique is very fast, requires little laboratory resources, and can replace rapid antigen tests or verify reactive rapid tests on-site.

Keywords: COVID-19; LAMP; RT-PCR; SARS-CoV-2; direct testing; loop-mediated isothermal amplification; nucleic acids.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 Nucleic Acid Testing / methods*
  • DNA, Viral / analysis*
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Nasopharynx / virology*
  • Nucleic Acid Amplification Techniques / methods*
  • SARS-CoV-2 / isolation & purification*
  • Sensitivity and Specificity
  • Specimen Handling

Substances

  • DNA, Viral

Supplementary concepts

  • LAMP assay