Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE

Aims: Development of a simple, specific, rapid and inexpensive Dot-ELISA te st for diagnosis of rotaviral antigen in stool samples. Methods and Results: Hyperimmune rabbit antisera raised against SA-11 (Simi an Agent-11) strain was used as primary antibody. The secondary antibody co njugate used was the goat anti-rabbit IgG alkaline phosphatase, and BCIP/NB T solution was used as substrate. Faecal extracts were diluted 10-fold and used for the detection of rotavirus antigen. RNA-PAGE was performed to comp are the specificity and sensitivity of the diagnostic tests. Dot-ELISA posi tive samples were further confirmed by Western blot analysis. Conclusions: This Dot-ELISA test could be used as an alternative method for diagnosing rotaviral samples in the field. Significance and Impact of the Study: The Dot-ELISA test is simple, specifi c, rapid and cost effective. It is suitable for identifying a large number of samples obtained from epidemiological studies and hence, reducing the de ath rate of rotavirus-infected patients.