Development of a rapid detection kit for chicken infectious anemia virus nucleic acid (RPA-LFD method).

Abstract

Chicken infectious anemia virus (CIAV) is a major immunosuppressive pathogen that causes huge economic losses in the poultry industry. The virus persists in the host for an extended period, making it difficult to eliminate and highlighting the need for rapid and accurate detection methods. In this study, we developed a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) kit for on-site detection of CIAV. Optimal primers and probes were designed and screened based on conserved regions of the full-gene sequences of 251 CIAV strains, ensuring high specificity, with no cross-reactions with 12 other common avian viruses. It demonstrated high sensitivity with a minimum detection limit of 10 copies of CIAV DNA, achieving efficient detection in just 15 min. The kit also showed good repeatability and stability, with 100 % concordance with qPCR in detecting 50 field samples. The developed CIAV RPA-LFD kit provides a practical tool for the rapid detection of CIAV, suitable for both on-site and laboratory use.