A reverse transcription recombinase-aided amplification assay for the rapid detection of goose astrovirus.

Abstract

China's burgeoning animal husbandry sector has witnessed a notable expansion in goose farming. Among the various health challenges, a novel goose astrovirus (GoAstV) has emerged as a significant threat to the industry, necessitating prompt detection strategies to mitigate its economic impact. This research introduces a novel detection approach using real-time fluorescence-based reverse transcription recombinase-aided amplification (RT-RAA), offering a rapid and reliable method for GoAstV identification. We meticulously designed specific primers and probes, and optimized the RT-RAA reaction conditions. The assay's specificity, sensitivity, repeatability, and clinical efficacy were rigorously assessed. Our method achieves detection within a swift 26-min window at a constant temperature of 39.00 ˚C, boasting a detection threshold as low as 1.19 × 10² copies per μL. Notably, the assay exhibited no cross-reactivity with closely related viruses, including Newcastle disease virus, avian influenza virus H9 subtype, goose circovirus, goose parvovirus, duck Tembusu virus, and avian adenovirus type 4. Validation through testing of 40 clinical samples confirmed a 100% agreement with pre-existing data. The study's outcomes underscore the high specificity, sensitivity, and operational simplicity of the developed RT-RAA assay, positioning it as an ideal candidate for the rapid and on-site detection of GoAstV.