An enzyme immunoassay for the qualitative and semiquantitative determination of IgA-class antibodies to Bordetella pertussis and Bordetella pertussis toxin in serum and plasma.
The DRG Bordetella pertussis / toxin IgA ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA). Microtiter wells as a solid phase are coated with Bordetella pertussis and Bordetella pertussis toxin antigen. Diluted patient specimens and ready-for-use controls are pipetted into these wells. During incubation Bordetella pertussis and Bordetella pertussis toxin-specific antibodies of positive specimens and controls are bound to the immobilized antigens. After a washing step to remove unbound sample and control material horseradish peroxidase conjugated anti-human IgA antibodies are dispensed into the wells. During a second incubation this anti_IgA conjugate binds specifically to IgA antibodies resulting in the formation of enzyme-linked immune complexes. After a second washing step to remove unbound conjugate the immune complexes formed (in case
of positive results) are detected by incubation with TMB substrate and development of a blue color. The blue color turns into yellow by stopping the enzymatic indicator reaction with sulfuric acid. The intensity of this color is directly proportional to the amount of Bordetella pertussis and Bordetella pertussis toxin-specific IgA antibody in the patient specimen. Absorbance at 450 nm is read using an ELISA microtiter plate reader.