Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE.
OBJECTIVE
Development of a simple, specific, rapid and inexpensive Dot-ELISA test for diagnosis of rotaviral antigen in stool samples.


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.


CONCLUSIONS
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.
Department of Microbiology, Osmania University, Hyderabad, India.
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T
Anand
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Narasa Raju
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C
Vishnu
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Venkateswar Rao
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Sharma
D000956:Antigens, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000737:chemistry; D006801:Humans; D012367:RNA, Viral / Q000032:analysis; D012401:Rotavirus / Q000737:chemistry* / Q000235:genetics
D000956:Antigens, Viral; D012367:RNA, Viral