Visual and Equipment-Minimized Detection of Avian Metapneumovirus Subtype C by RT-RPA-CRISPR/Cas12a Coupled with Lateral Flow Dipstick

Abstract

Simple Summary Avian metapneumovirus, particularly the C type, causes severe respiratory disease and egg drop in ducks, leading to significant economic losses in the poultry industry. Current diagnostic methods rely on laboratory-based tests that require expensive equipment and take several hours, making them impractical for use on farms. To address this problem, we developed a rapid, equipment-minimizing detection method that combines a fast nucleic acid amplification technique with a gene-targeting system and a simple paper strip for visual result reading. Using samples collected from ducks, the method produced results within one hour using only a basic heating device, and a colored line on the strip indicated a positive infection. When compared with the most reliable laboratory test currently available, our method showed 98.85% agreement. This approach offers a practical option for near-farm testing and has the potential to help farmers and veterinarians quickly identify infections, enabling timely intervention and reducing economic losses. Future improvements, such as dry-format reagents that do not require cold storage, will further advance its use in field settings.