Description
Principle of the assay: Bordetella pertussis IgG ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-Bordetella pertussis IgG antibody and an Bordetella pertussis IgG-HRP conjugate. The assay sample and buffer are incubated together with Bordetella pertussis IgG-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the Bordetella pertussis IgG concentration since Bordetella pertussis IgG from samples and Bordetella pertussis IgG-HRP conjugate compete for the anti-Bordetella pertussis IgG antibody binding site. Since the number of sites is limited, as more sites are occupied by Bordetella pertussis IgG from the sample, fewer sites are left to bind Bordetella pertussis IgG-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Bordetella pertussis IgG concentration in each sample is interpolated from this standard curve