Abstract: The aim of this study was to obtain Canine Parvovirus (CPV) strains in the middle of China and study their genetype, biological characterization and pathogenicity. 14 feces swabs that were positive by colloidal gold test were collected from pet clinics in Wuhan. Then they were passaged in Feline kidney 81 (F81). 2 pairs primers were designed according to conservative Capsid protein 2 (VP2). The strains were purified by plaque assay, and then onestep growth curves were obtained. Subsequently transmission electron microscopic (TEN) sections were made. And the animal experiment was carried out. 5 CPV strains were isolated. 390 bp and 583 bp nucleotide sequences were acquired. After sequencing and analyzing the gene fragments, the result shows that 4 strains were CPV2a and 1 was CPV2b. The curves show the growth of CPV at different stages after infection. The reproductive activity can reach or beyond 105.5 PFU·mL-1. The photographs of TEN showed that mitochondria are swelling. Finally, animal experiment shows that every dog had clinical symptoms in different degrees. The results of dissection reflect that there are necrosis in the small intestine and the myocardium becomes thinner. After pathological sections were made, observation was taken up and the result shows that myocardial cells are metamorphic and necrotic. This study provides groundwork for further study of molecular biology and pathopoiesia of CPV.