A sensitive and simple RT-LAMP assay for sarbecovirus screening in bats

Microbiol Spectr. 2023 Dec 12;11(6):e0259123. doi: 10.1128/spectrum.02591-23. Epub 2023 Nov 16.

Abstract

We report the application of a colorimetric and fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to facilitate mass screening for sarbecoviruses in bats. The assay was evaluated using a total of 838 oral and alimentary samples from bats and demonstrated comparable sensitivity and specificity to quantitative reverse transcription PCR (qRT-PCR), with a simple setup. The addition of SYTO9, a fluorescent nucleic acid stain, also allows for quantitative analysis. The scalability and simplicity of the assay are believed to contribute to improving preparedness for detecting emerging coronaviruses by applying it to field studies and surveillance.

Keywords: RT-LAMP; SYTO9; bat screening; coronavirus; molecular detection; sarbecovirus.

MeSH terms

  • Animals
  • Chiroptera* / virology
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • Reverse Transcription
  • Severe acute respiratory syndrome-related coronavirus*

Supplementary concepts

  • LAMP assay