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Test Principle: The ELISA is based on the the qualitative enzyme immunoassay technique.The Microplate provided in this kit has been pre-coated with an Antibody specific to E.coli O157, make it to solid-phase antibody.Samples are added to the Microplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)-conjugated antibody specific for E.coli O157 is added to each Microplate well and incubated,so the antibody-antigen-Enzyme labeled antibody complex is formed.Following a wash to remove any unbound reagent,then the TMB substrate solution is added to each well. Only those wells that contain E.coli O157 and HRP conjugated E.coli O157 antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The qualitative determination of E.coli O157 is determined by comparing with the CUTOFF value.