The structure of florfenicol was transformed to prepare the hapten antigen and antigen, the polyclonal antibody was obtained by immunized rabbit.The determination of the antibody showed high sensitivity and have reactions on florfenicol and florfenicol amine, the titer was 1∶480000. On this basis, the indirect competitive ELISA method was established through the optimization of the parameters.The detection limit of the established ELISA method was 0.5μg/kg, the sample recovery was 82.5% to 96.0% by 20,50μg / kg addition to the blank experiment, and the inter-assay coefficient ranged from 4.1% to 15.2%. The method could be used for preliminary detection of florfenicol residues in animal tissue samples。