Objective To establish an animal model of positive antisperm antibody (AsAb) in rats. Methods The animal model was set up by artificial immunity with iso-Wistar rat sperm antigen. The level of AsAb in serum was determined by ELISA. The morphological changes of the epididymis tissue in rats were determined by optical microscope with HE. The rats sperm was visualized by staining acrosome with the dye Coomassie brilliant blue. The changes of ATPase activity of the rats sperm were detected by chemical colorimetry. Results The AsAb of rats serum in the model accorded with the experimental demand. The percentages of acrosome reaction, Na -K ATPase activity, Ca 2-ATPase activity in AsAb positive rats sperm significantly decreased as compared with the control group (P<0.01; P<0.05; P<0.05).Conclusion The results demonstrated it was successful to establish an animal model of AsAb positive rats.