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IDENTIFICATION AND DETECTION OF WATER-BORNE VIRUSES BY IMMUNOENZYMATIC METHODS
Citation:
Herrmann, J. IDENTIFICATION AND DETECTION OF WATER-BORNE VIRUSES BY IMMUNOENZYMATIC METHODS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/4-80/034.
Description:
A quantitative enzyme-linked immunosorbent assay (ELISA) was used for identification of viruses selected as representative water-borne viruses: poliovirus 1, echovirus 6, coxsackievirus A9, and coxsackie B viruses. Partially purified viral antigens or virus-specific antibodies were absorbed to polystyrene spectrophotometer cuvettes, which permitted the assays to be reported and compared in terms of enzyme units specifically reacting. Inhibitors in diluents used to prevent non-specific adsoption of immunoreagents caused desorption varied with the type of preparation used, and antibody desorption was dependent on the concentration of antibody initially adsorbed. For specific identification of a given enterovirus type by the ELISA method, approximately 100,000 plaque-forming units of virus per assay tube were required. To alleviate the problem of antibody and virus desorption, antibodies and virus were immobilized by covalent linkage on nylon balls for use in solid-phase enzyme-linked immunoassays. A higher percentage of virus could be immobilized by this method than was possible by adsorption to polystyrene, and enzyme-linked immunoassay on nylon was sufficiently specific to differentiate the three poliovirus types.