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

Area of Science:

• Virology
• Molecular Biology
• Public Health

Background:

• Coronaviruses, particularly sarbecoviruses, pose a significant threat for zoonotic spillover.
• Effective surveillance in bat populations is crucial for early detection of emerging viral threats.
• Existing diagnostic methods can be resource-intensive for large-scale field screening.

Purpose of the Study:

• To develop and evaluate a novel colorimetric and fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for mass screening of sarbecoviruses in bats.
• To assess the assay's performance against established quantitative reverse transcription PCR (qRT-PCR) methods.
• To highlight the assay's utility for field-based surveillance of bat-borne coronaviruses.

Main Methods:

• Development of a colorimetric and fluorescent RT-LAMP assay.
• Evaluation of the assay using 838 oral and alimentary samples from bats.
• Comparison of RT-LAMP assay sensitivity and specificity with qRT-PCR.
• Inclusion of SYTO9 fluorescent nucleic acid stain for quantitative analysis.

Main Results:

• The RT-LAMP assay demonstrated comparable sensitivity and specificity to qRT-PCR.
• The assay provided a simple and scalable method for detecting sarbecoviruses.
• Quantitative analysis was achievable with the addition of SYTO9.

Conclusions:

• The developed RT-LAMP assay is a valuable tool for mass screening of sarbecoviruses in bats.
• Its simplicity and scalability enhance preparedness for detecting emerging coronaviruses.
• The assay is suitable for field studies and ongoing surveillance programs.