Abstract: Objective This study was aimed at developing rapid detection by PCR for leptospire in order to apply this rapid detection on host animals and comparing the effectiveness of the detection by PCR and by convention method. Methods A rapid detection for leptospire by PCR was established with the primers from Chinese Center for Disease Control and Prevention. Enteropathogenic E.coli, Enterotoxigenic E.coli, Vibrio alginolyticus, Saimonella, Cholera bacillus, Shigella dysenteriae, Cholera bacillus of non-O1, O139, Vibrio parahaemolyticus and Enterohemorrhagic E. coli O157:H7 were used as control to detect leptospire rapidly. Results The PCR results of detection for leptospiral strains isolated in Quzhou of 2006 were consisitent with the serological identification. Specimens of liver, spleen and kidney from 100 mice and 100 frogs were detected by PCR, with the positive rate being 10% while the isolation rate was only 1%(2=16.05, P0.005). There was statistical difference between the two groups of data. Conclusion The high sensibility and powerful specificity of the PCR detection made it suitably applied in the detection of leptospire.