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Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE
T Anand1, T A Narasa Raju, C Vishnu
1Department of Microbiology, Osmania University, Hyderabad, India.
Aims:
Development of a simple, specific, rapid and inexpensive Dot-ELISA test for diagnosis of rotaviral antigen in stool samples.
Methods And Results:
Hyperimmune rabbit antisera raised against SA-11 (Simian Agent-11) strain was used as primary antibody. The secondary antibody conjugate used was the goat anti-rabbit IgG alkaline phosphatase, and BCIP/NBT solution was used as substrate. Faecal extracts were diluted 10-fold and used for the detection of rotavirus antigen. RNA-PAGE was performed to compare the specificity and sensitivity of the diagnostic tests. Dot-ELISA positive 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, specific, rapid and cost effective. It is suitable for identifying a large number of samples obtained from epidemiological studies and hence, reducing the death rate of rotavirus-infected patients.