Immunoglobulin class and subclass-specific monoclonal antibody sandwich ELISA for the detection of antibodies against coxsackieviruses B, types 1-5.

Immunoglobulin subclass-specific ELISAs were developed for human IgG1, IgG2, IgG3, IgG4, IgAtotal, and IgM directed against Coxsackie B (CB) virus types 1, 2, 3, 4, and 5. In all the assays the solid phase was coated with immunoglobulin class/subclass-specific monoclonal antibodies, followed by an incubation with the serum specimens. Incubation with one of the CB viruses, as well as an incubation with biotinylated serotype-specific monoclonal antibodies to the same virus type provided the virus specificity. Finally, there were incubations with peroxidase labeled Extravidin and the substrate-chromogen system. This ELISA method eliminated the competition between the immunoglobulin classes and subclasses. IgG3 and/or IgG1 were seen most frequently of the IgG subclasses, but IgG2 and IgG4 were also present infrequently. The viral specificity of the antibody subclass assays seems to be predominantly at the enterovirus group level, but this remains to be evaluated in a larger study. IgA and IgM were seen almost exclusively in specimens from patients with acute enteroviral infections, except in the assays with the crude CB5 antigen. This indicates the possible suitability of the IgA and IgM assays as diagnostic tests for enteroviral infections. A larger study is necessary to confirm this finding.