The quantitative detection of zearalenone in feed by enzyme-linked immunosorbent assay (ELISA) is based on competitive enzyme-linked immunosorbent assay (ELISA). Extract zearalenone from crushed samples by shaking with methanol/water, filter the aqueous extract, and then perform immunological testing. Add the enzyme label of zearalenone to the mixed well with standard or sample. After mixing the sample enzyme label, 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 zearalenone 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 the stop solution and read the absorbance value. Compare the unknown concentration sample with the standard absorbance value to obtain the concentration of zearalenone in the sample.