Adenovirus ELISA

The Adenovirus ELISA employs monoclonal antibodies in a sandwich-type method. A monoclonal antibody to the hexon antigen of the adenoviruses is coated to the surface of each well of the microplate. 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 monoclonal anti-adenovirus antibodies (Conjugate 1) for incubation at room temperature (20 - 25° C). After washing, streptavidin poly-peroxidase conjugate (Conjugate 2) is added and incubated again at room temperature (20 - 25° C). With the presence of adenoviruses in a stool sample, a sandwich complex forms which consists of immobilized antibodies, the adenovirus antigens, and the antibodies conjugated with the biotin-streptavidin-peroxidase complex. Unattached streptavidin poly-peroxidase conjugate is removed in a subsequent washing step. After addition of the substrate, the attached enzyme changes the previously colorless solution to blue in the wells that contain bound toxins. Upon addition of the stop solution, the color changes from blue to yellow. The absorbance is proportional to the concentration of adenoviruses present in the sample.