Solid phase indirect radioimmunoassay: Standardization and applications in viral serology

Abstract

In solid phase indirect radioimmunoassay (IRIA) antiviral immunoglobulins (Ig) bind specifically to the viral antigen fixed onto wells of microtiter plates. Radioactively labeled (125I) anti-Ig can be used to detect these antiviral antibodies. The sensitivity of the IRIA depends on the amount of antigen in the microtiter wells and the concentration of the 125I-labeled anti-Ig used. To standardize the assay, a method of antigen titration in the IRIA was devised, using parainfluenza type 1 viruses as antigens. The IRIA provides a means to titrate viral antigens with different biologic activities and antibodies against them. A binding inhibition test (BIT) based on IRIA allows the antigenic analysis of different virus strains, as is demonstrated for two closely related parainfluenza type 1 virus strains.

Immune Responses to Avian Influenza Viruses

“…Severe lymphopenia is a hallmark of severe infection with avian IAVs in humans and in mice (65)(66)(67). Two H5N1 viruses were selected for evaluation in a mouse model.…”

Serological Methods in the Identification and Characterization of Viruses

“…The reader is referred to that review, which covers advances up to 1976. Subsequent work has been concerned mainly with the development of additional solid-phase procedures that meet the requirements of speed, sensitivity, and large-scale operation (Middleton et al, 1977;Yung et al, 1977). Using an indirect radioimmunoassay with 1251-labeled rabbit antibovine IgG for detecting rotavirus antibody, Babiuk et al (1977) found that the assay was a thousandfold more sensitive than the counterimmunoelectrophoresis technique.…”

Development, comparison and utilization of enhanced immunoassays for the detection of lettuce mosaic virus

“…Furthermore, spontaneous elution of antibody is more likely to take place at high concentrations of coating antibody (Engvall and Perlman, 1971;Hermann and Collins, 1976;Yung et al, 1977). In addition, higher concentrations of coating antibody give higher non-specific reactions.…”