The detection of vomitoxin in feed by enzyme-linked immunosorbent assay (ELISA) is based on competitive enzyme-linked immunosorbent assay (ELISA). Shake and extract DON from the crushed sample with water, filter the aqueous extract, and then perform immunological testing. Add the enzyme label of vomitoxin to the test well that has been added with standard or sample. After the antibody is added, the reaction begins, and the sample and enzyme label compete to bind to the antibody attached to the micropore. After 10 minutes of cultivation, pour out the solution from the well and wash away the unbound vomitoxin and enzyme label from the micropore. Add colorless substrate solution and incubate for 5 minutes. All bound enzyme markers convert the substrate into a blue substance. Add stop solution and read the absorbance value. Compare the unknown concentration sample with the standard absorbance value to obtain the concentration of vomitoxin in the sample.