Establishment and Preliminary Application of an ELISA Method for Detecting Antibodies to Field Strains of Bovine Infectious Rhinotracheitis Virus.

BICP0-ELISA enables accurate IBRV detection and differentiates infected from vaccinated cattle.

This study describes the development and validation of an indirect ELISA (BICP0-ELISA) for detecting antibodies to bovine infectious rhinotracheitis virus (IBRV) in cattle. The assay targets epitope 3 of the BICP0 non-structural protein, which is expressed only during active IBRV infection, not following inactivated vaccination. Recombinant BICP0 protein was expressed in E. coli using codon-optimized sequences and purified for use as coating antigen. Optimal assay parameters were established through systematic optimization. ROC curve analysis determined a cut-off value of 0.225, achieving 100% sensitivity and 98.6% specificity when compared to a commercial ELISA kit. Field evaluation of 274 serum samples from cattle farms in Liaoning Province revealed overall IBRV seropositivity of 40.1% (28.5% in non-vaccinated herds, 50.9% in vaccinated herds). The BICP0-ELISA demonstrates excellent analytical performance and importantly provides DIVA (differentiate infected from vaccinated) capability in herds vaccinated with inactivated vaccines, supporting its utility for IBR surveillance and eradication programs.

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