Development of a monoclonal antibody-based sandwich ELISA for detection of equine rotavirus from diarrhoeic foals

A sensitive and specific sandwich enzyme-linked immunosorbent assay (ELISA) was developed employing a monoclonal antibody (mAb) raised against group-specific protein VP6 of rotavirus, for detection of equine rotavirus (ERV) from stool samples. This assay specifically detected rotavirus and did not react with other representative equine viruses. On testing of stool samples from diarrhoeic foals (n=72), 23 were detected positive by ELISA, whereas RNA-PAGE could detect only 11 samples positive. The ELISA was found 100% sensitive while the sensitivity of RNA-PAGE was only 52.38% in comparison to virus isolation. The ELISA was also very specific, giving a specificity of 0.96 (0.8541≤π≤0.9932, p<0.05) in comparison to virus isolation. Results obtained by both virus isolation and ELISA (= 0.9346) were comparable. The prevalence of rotavirus in diarrhoeic foals below two months of age in an organized farm located in northern India was nearly 32% as detected by this ELISA. The mAb-based ELISA developed in the present study is simple to perform, highly sensitive and specific assay for detection of rotavirus from equine stool samples.