Use of the nitrocellulose-enzyme immunosorbent assay for rapid, sensitive and quantitative detection of human enteroviruses

Abstract

A modified enzyme immunosorbent assay (EIA) employing nitrocellulose (NC) membrane as a high-capacity solid phase was successfully employed for the sensitive and rapid detection of human enteric viruses, poliovirus and Coxsackievirus B-5. The sensitivity of the NC-EIA ranged from 7 to 70 pg of viral antigen diluted in phosphate-buffered saline. When virus was added to crude supernatants of mollusc tissue homogenates prepared by the standard procedure for the recovery of viruses in molluscs, the sensitivity was reduced by approximately 10-fold. The sensitivity of the NC-EIA for the detection of viral antigens was 10- to 100-fold higher than conventional EIA using polystyrene microtiter plates as solid phase supports. This simple, rapid and sensitive assay using minimal amounts of antibodies should prove to be a useful and practical diagnostic tool to augment infectivity assays currently employed by various virus monitoring procedures. The method also may be applicable for the detection of difficult to grow and/or non-cultivatable enteric viruses which may be present in sewage-contaminated environments.