Abstract: In this study, a direct competitive biomimetic enzyme-linked immunosorbent assay (BELISA) method using molecularly imprinted polymers (MIPs) as a bionic antibody was developed to detect enrofloxacin (ENRO) in animal food. The MIPs were characterized by scanning electron micrographs (SEM), fourier transform infrared spectrometer (FT-IR), adsorption kinetics experiment and isothermal adsorptions experiment, respectively. Results showed that molecularly imprinted polymers were successfully synthesized with a fast adsorption rate and high adsorption. Under the optimum conditions, the sensitivity (IC50) and limit of detection (IC15) of the established BELISA method were 85.7 ng/mL and 1.1 ng/mL, respectively. The method was applied to the determination of ENRO in spiked beef and shrimp samples, and recoveries ranged from 74.4 to 86.8% with relative standard deviations (for n=3) between 3.3 and 8.0 %. Results correlated well with that obtained using ELISA and high performance liquid chromatography method