Abstract: As a new type of molecular recognition element, aptamer has the advantages of being able to be synthesized in vitro, easy to modify, and good stability. It is also considered as a good substitute for antibodies. Since the zearalenone aptamer was successfully screened, based on the high affinity and specific binding of the aptamer to the target, an enzyme-linked aptamer(ELAA)indirect competition method was developed for rapid detection of zearalenone (ZEA). Through the combination of biotin and streptavidin, aptamer was immobilized on the microplate 1; DNA1 was a partially complementary strand of the aptamer labeled with horseradish peroxidase (HRP); DNA2 was completely complementary to DNA1 single-stranded and fixed to the microplate 2 by biotin and avidin. When the sample has no ZEA, DNA1 and aptamer are partially complementary to each other and then fixed on the microplate 1. After adding TMB to the microplate 2, the color will not develop; when the sample contains ZEA, the target will specifically bind to the aptamer, making DNA1 melt in a free state. DNA1 solution was pipetted to the microplate 2, the DNA1 labeled with HRP is captured by the DNA2 in the microplate 2 based on the principle of double-strand complementary pairing, catalyzing the TMB substrate turn to blue, and H₂SO₄ was added to stop the reaction and the color will turn to yellow. The absorbance was detected at the wavelength of 450 nm. Therefore, the higher the ZEA content, the more it binds to the ZEA aptamer, and the more free DNA1, that is, the more DNA1 captured on the microplate 2, and the deeper the color of the catalytic TMB and the greater the absorbance value. The result of the enzyme-catalyzed color reaction was used for ZEA detection. Key parameters, such as streptavidin, blocking solution, aptamer and complementary chain concentration were optimized. The optimized conditions were as follows:dry coating method was used; SA was 4 μg/mL; mass fraction of BSA was 2%; aptamer was 270 nmol/L; Bio-DNA1 was 10 nmol/L; Bio-DNA2 was 90 nmol/L. Moreover, the sensitivity, specificity and practicality of the assay were also studied. Under the optimal conditions, a standard curve was establish with the regression equation of y=0.005 9x+2.497 6 (R²=0.995 9), the mass concentration of ZEA and the absorbance of the reaction system showed good linearity in the range of 0.5 to 100 ng/mL with the lowest detection limit of 0.44 ng/mL. In addition, the developed measurement method was used for specific analysis and detection of several mycotoxins, such as AFB₁, OTA, and DON, which often coexist with ZEA. Finally, ELAA has good specificity for corn and corn oil. The results were consistent with the ELISA results. This research will provide a simple, effective and economical method to quickly detect ZEA in cereal products.