Campylobacter ELISA

The Campylobacter ELISA employs monoclonal antibodies in a sandwich-type method. The well surface of the microwell plate is coated with monoclonal antibodies to Campylobacter antigens. A pipette is used to place a suspension of the stool sample to be examined, as well as control samples into the wells of the microplate together with biotinylated anti-Campylobacter antibodies (Conjugate 1) for incubation at room temperature (20 – 25 °C). After a wash step, streptavidin poly-peroxidase conjugate (Conjugate 2) is added, and it is incubated again at room temperature (20 – 25 °C). With the presence of Campylobacter antigens in a sample, immobilized antibodies, antigens, and conjugated antibodies form a sandwich complex. Another wash step removes the unattached streptavidin poly-peroxidase conjugate. After adding the substrate, the attached enzyme changes the color of the previously colorless solution in the wells of the microplate to blue if the test is positive. Addition of a stop reagent changes the color from blue to yellow. The signal observed is proportional to the concentration of the Campylobacter antigen that is found in the sample.