Enzyme-linked immunosorbent assay for human IgG, IgA, and IgM antibodies to antigens from anaerobic cultures of seven oral bacteria.
Monocultures of 7 oral bacteria were grown anaerobically, and antigens were partially purified from the supernatant fluids by gel filtration. Isoelectric focussing showed that the antigen fractions contained PAS-positive material and proteins focussing between pH 3.5 and pH 5.5. Specific activity against the 7 antigen fractions was observed for IgG, IgA, and IgM in serum from patients with periodontal disease by means of enzyme-linked immunosorbent assay (ELISA). Conditions for antigen coating in ELISA were studied, and large variations with regard to optimal concentration were found among the 7 fractions. The ELISA readings were virtually unaffected by increasing the pH of the coating buffer from 5 to 9. Addition of sheep serum to the incubation buffer was disadvantageous as the serum showed moderate antibody activity to the 7 antigen fractions, and human serum apparently contained IgM antibodies to sheep IgG. Addition of 0.5% BSA improved the reproducibility of ELISA considerably, although the sensitivity was somewhat decreased.
D000693:Anaerobiosis; D000818:Animals; D000907:Antibodies, Bacterial; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial; D001419:Bacteria / Q000254:growth & development*; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005673:Fusobacterium / Q000276:immunology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011817:Rabbits; D012756:Sheep; D014678:Veillonella / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M