Objective To establish special pathogen free Kunming mouse of Hubei closed colony.Method The purification mouse was made by excising operation of uterus in the situation of bacteria free. The SPF Kunming mouse strain of Hubei closed colony was developed with the following surveillance on microbes and parasites in first generation.Results 57 mice were got from 67 cases by operation, and 509 baby mice with purification,from which 460 grew into adults and the state of microbiology in those mice was in keeping SPF.Conclusion The method can keep the excellent laboratory animal stain with biological purification.