Simplified assay for measuring Toxoplasma gondii immunoglobulin G avidity.
A Toxoplasma gondii immunoglobulin G (IgG) avidity enzyme-linked immunosorbent assay (ELISA) was developed that combines the accuracy of assays based on end point titers and the relative ease of assays based on optical density values. Like published procedures, the new assay's avidity index (AI) was based on differential T. gondii-specific IgG reactivity in serum-treated wells washed with urea buffer versus that in wells washed with control buffer; unlike previous assays, however, the IgG reactivity was measured quantitatively using a standard curve. The assay was evaluated using 24 IgG-positive and IgM-positive sera collected within 5 months of the onset of symptoms (recent-infection group) and 25 IgG-positive and IgM-negative sera (past-infection group). All sera in the recent-infection group exhibited AI values of <0.18, whereas all sera in the past-infection group exhibited AI values of >0.27. The AI values of the recent-infection group showed significant correlation with the number of days after the onset of symptoms. A subset of 16 sera (8 recent and 8 past) was tested using a commercially available T. gondii IgG avidity ELISA based on end point titration; the results of the two assays showed highly significant correlation (R(2) = 0.9125). In addition, we confirmed and extended the findings of other investigators, showing that AI values calculated using optical density values, but not AI values calculated using quantitative IgG values, varied significantly depending on the serum dilution used. This new assay should facilitate the accurate measurement of T. gondii IgG avidity in a reference laboratory setting.
Prince
H E
HE
Focus Technologies, 5785 Corporate Ave., Cypress, CA 90630, USA. hprince@focusanswers.com
Wilson
M
M
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood / Q000378:metabolism; D000915:Antibody Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000378:metabolism*; D007075:Immunoglobulin M / Q000097:blood / Q000378:metabolism*; D011933:Reagent Kits, Diagnostic / Q000469:parasitology; D012016:Reference Values; D014122:Toxoplasma / Q000276:immunology*; D014123:Toxoplasmosis / Q000097:blood / Q000175:diagnosis
D000913:Antibodies, Protozoan; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011933:Reagent Kits, Diagnostic