Development and evaluation of an enzyme-linked immunosorbent assay for measuring antibodies against VP8∗ receptor binding domain of equine rotavirus A.

Abstract

Equine rotavirus A (ERVA) poses a significant threat to foal health, necessitating reliable serological tools for assessing protective immune responses. This study focused on the development of an enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies against the VP8∗ receptor binding core domain of ERVA. Following the expression and purification of VP8∗ receptor binding domain from the bacterial expression system, the ELISA for detecting anti-ERVA VP8∗ antibodies with VP8∗ as a coating antigen was developed and its performance was validated by the comparative analysis with a virus neutralization assay (VNA). Our results showed a good correlation between ELISA and VNA in evaluating protective antibody responses to ERVA. The VP8∗-based ELISA demonstrated good sensitivity and specificity, effectively distinguishing vaccinated from non-vaccinated horses. Finally, we observed a strong correlation between ELISA and virus neutralization assays, further supporting its utility as a simple and cost-effective approach for measuring protective antibodies against ERVA.