Description
Break-apart microwells are coated with C. difficile GDH specific monoclonal antibodies. A horseradish peroxidase (HRP) conjugated monoclonal anti-GDH antibody is added to the antibody-coated microwells. Fecal samples are diluted in sample diluent and added to the microwells. In this step C. difficile GDH is marked by the HRP conjugated antibody and immobilized by the coated antibody. Unbound conjugate is removed by washing. TMB-substrate is added, the substrate is hydrolyzed by the peroxidase and yields a blue solution of the reduced substrate. Upon the addition of the stop solution, the blue color turns yellow and should be read by an ELISA reader at a wavelength of 450/620 nm. The absorbance is proportional to the level of C. difficile GDH in the sample