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Development of Dot‐ELISA for the detection of human rotavirus antigen and comparison with RNA‐PAGE
Author(s) -
Anand T.,
Raju T.A. Narasa,
Vishnu C.,
Rao L. Venkateswar,
Sharma G.
Publication year - 2001
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2001.00884.x
Subject(s) - rotavirus , virology , antigen , rna , biology , reoviridae , microbiology and biotechnology , virus , gene , immunology , genetics
T. ANAND, T.A. NARASA RAJU, C. VISHNU, L. VENKATESWAR RAO AND G. SHARMA. 2001.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.