Objective To investigate the culture positive rate of people and mouse with leptospirosis in Qingyuan from 2000 to 2003. Methods Leptospira were cultured from the serum samples of patients and animals kidney. Anti leptospira antibodies were detected by the MAT method from the serum samples of health people, patients and mice. Results Leptospira were cultured from 17 samples of patientssera and animals kidney. The cultured leptospira were belong to Canicola, Autumnalis, Sejroe and Javanica sero groups. The positive rates of anti leptospira antibody of health people, patients and mice were 29.22%, 3.81%and 29.71%, respectively. Conclusions The positive rates of anti leptospira antibody of healthy people and mice were high. It was the first time that the Sejroe sero group of leptospira cultured from patient serum samples. Surveillance of leptospira should be enhanced. A more comprehensive control measures should be taken to prevent and control of leptospirosis.