Chemiluminescent immunoassays: Discrimination between the reactivities of natural and human patient antibodies with antigens from eukaryotic pathogens, Trypanosoma cruzi and Paracoccidioides brasiliensis

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Abstract
Quantitative chemiluminescent enzymelinked immunosorbent assay (ELISA) and dot-blotting procedures were developed to evaluate the reactivity of human antibodies with crude antigens and purified molecules of parasites and fungi, mainly Trypanosoma cruzi and Paracoccidioides brasiliensis. Reproducible, highly sensitive, and strictly doseresponding results were obtained, with the specificity depending on the kind of antigen used. Mixed antigens (epimastigote membrane and HIV-1 heptapeptide) applied in dots could be independently recognized by specific sera. Purified antigens (T. cruzi F2/3 and P. brasiliensis gp43) at very small concentrations gave specific reactions with patients' sera diluted ≥1:1,000 and were very poorly reactive or unreactive with natural antibodies using the chemiluminescent immunoassays. P. brasiliensis crude antigen Fava Netto polysaccharide antigen (FNPA) contained peptide epitopes recognized by natural antibodies and carbohydrate epitopes reactive with sera from histoplasmosis patients. It is very important that sensitive chemiluminescence immunoassays be used with purified antigenic molecules to ensure specificity for the diagnosis and follow-up of parasitic and fungal infections.

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