Limitations of the radioimmunoprecipitation polyethylene glycol assay (RIPEGA) for detection of filarial antigens in serum.
The performance of the radioimmunoprecipitation polyethylene glycol assay (RIPEGA) was examined for quantitation of filarial antigens (Brugia malayi and Dirofilaria immitis) in serum from infected human and animal hosts and non-infected controls. Multiple PEG concentrations were employed to determine the level of non-specific binding (NSB) in non-exposed human sera (NEHS) containing no filarial antigen. The NSB observed when 3 different 125I-labeled IgG antibodies were added to 26 NEHS varied 3-fold and was correlated significantly with total serum IgM (r = 0.80, P less than 0.005, n = 24) but not with serum IgA (r = 0.37) or IgG (r = 0.45). NSB levels were significantly reduced when a Fab'2 fragment of the 125I-labeled antibody was used, but the correlation of NSB with total serum IgM remained significant (r = 0.57, P less than 0.01). The presence of rheumatoid factor in NEHS sera also significantly increased NSB by an average of 3-fold. These effects eliminated the assay's ability to detect in sera from infected hosts filarial antigen the presence of which could be readily demonstrated by an immunoradiometric assay. The RIPEGA's precision (intra-assay coefficient of variation (CV) = 21% at 35% Bmax) and reproducibility (inter-assay CV = 29% at 35% Bmax) are less satisfactory than many alternative immunoassays. In many cases, positive sera failed to dilute out in parallel with each other or with an antigen-spiked standard reference curve. We conclude that poor performance characteristics currently limit the utility of the RIPEGA for quantitating filarial antigen in human and animal serum.
D000818:Animals; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis*; D002009:Brugia / Q000276:immunology; D011232:Chemical Precipitation; D003240:Connective Tissue Diseases / Q000276:immunology; D004183:Dirofilaria immitis / Q000276:immunology; D004285:Dogs; D005368:Filariasis / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins / Q000502:physiology; D011092:Polyethylene Glycols; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D012217:Rheumatoid Factor / Q000502:physiology
D000941:Antigens; D007136:Immunoglobulins; D011092:Polyethylene Glycols; D012217:Rheumatoid Factor