Objective To establish a polymerase chain reaction method for canine parvovirus dection and to utilize the PCR for diagnosis of parvovirus in canine samples. Methods PCR primers were designed according to the .VP2 Gene of CPV. 221bp products was obtained with the template from CPV strain, while no band was found in infectious canine hepatitis virus. This amplified product was cloned into pMD18-T vector and transferred into E. coli JMI09. The recombinant plasmid was sequenced. 30 feces and 12 tissue samples were detected of CPV by PCR method. Results The identity of DNA sequence of this fragment between CPV strain and gi54646342/gi39748709/~57903634 was 100%, All of 30 feces samples was negative, 221bp fragment was amplified fronl 6 of 12 tissue samples, Conclusion A PCR method is established for the detection of parvovirus. The PCR assay is an sensitive and specific technique for parvovirus diagnosis.