Simplified radioimmunoassay for diagnostic serology

A simplified, indirect radioimmunoassay is described for Escherichia coli, vaccinia virus, and herpesvirus. The antigens were affixed to glass cover slips; thus both the primary and secondary reactions take place on the cover slips, and the unbound antiserum is easily separated from the bound antiserum by rinsing. Rabbit or human immune sera were reacted with the antigens, and the primary immune complex was quantitated by a secondary reaction with (125)I-indicator globulin (anti-rabbit or anti-human). A direct relationship between the antiserum concentration and the (125)I absorption was established. Variations in titers were detectable, and the titers were comparable to complement fixation titers. Homologous and heterologous reactions were distinguishable. The method affords an objective, quantitative, and qualitative evaluation of antibody, and results are reproducible.