LAUSR

Simple Summary Getah virus (GETV) is a zoonotic virus transmitted by mosquitoes. Various mammals are naturally infected with GETV, including foxes, pigs, horses, cattle, and sheep. To develop and validate a rapid, sensitive, and portable one-tube assay for the detection of GETV, we employed reverse transcription loop-mediated isothermal amplification (RT-LAMP) in conjunction with Pyrococcus furiosus Argonaute (PfAgo) to provide a straightforward and precise method for detecting porcine GETV in this study. LAMP is used to specifically amplify the target gene of GETV, after which PfAgo is directed to cleave the target gene and the probe, releasing a fluorescent signal. Additionally, the assay results can be observed under UV light or quantified using a fluorescence quantitative instrument. Our experiments demonstrate that this assay is highly specific, capable of detecting extremely low levels of the virus without cross-reacting with other viruses. When tested on real clinical samples, our novel method performed comparably to the gold standard quantitative real-time PCR test. This study provides a rapid, accurate, efficient, portable, and low-cost nucleic acid diagnostic method for the detection of GETV, which could serve as a powerful tool for the prevention and control of GETV.