Nine strains of canine parvovirus (CPV) were isolated using F81 cell from CPV positive feces collected from sick dogs in Changsha (CS)，the CPV strains were confirmed by PCR and hemagglutination assay showed all the virus isolates could agglutinate red blood cell of pigs with the titers all above 27. To further identify molecular and biological characteristics and the antigen variation of these isolates, VP2 gene was cloned, sequenced and analyzed. The results showed that the identities of gene sequence and the deduced amino acid sequence of CPV between the isolated strains and other reference strains were above 97% and 94%, respectively. And there were specific amino acid mutations in the VP2 of canine parvovirus CS strains.