Abstract
Bovine alphaherpesvirus 1 (BoHV-1) represents a significant threat to the cattle industry, emphasizing the need for reliable diagnostic tools that facilitate effective disease management. Current diagnostic methods, including virus neutralization tests (VNTs), are often complex and labor intensive, but existing antibody detection assays may lack sufficient accuracy. In this study, we developed a novel indirect enzyme-linked immunosorbent assay (iELISA) utilizing both BICP0 and gB proteins to enhance the detection of BoHV-1 infections. The inclusion of BICP0, a pivotal protein during the early stages of viral infection, markedly improved the assay's specificity and sensitivity. The BICP0-gB iELISA exhibited a high degree of concordance with the VNT, demonstrating superior sensitivity and specificity. Preliminary clinical evaluations revealed a real prevalence of 40.3 % (95 % CI: 33.1 %-48.0 %) in serum samples from yaks in Qinghai Province, which aligns with the documented prevalence of BoHV-1 in the region. These results underscore that BICP0-gB iELISA is a robust and reliable diagnostic tool for the differential diagnosis of BoHV-1, providing a cost-effective and efficient solution for high-throughput screening in the livestock industry.