Abstract: In order to understand the genetic variation characteristics of canine Parvovirus（CPV） in southern Henan Province, PCR method was used to amplify the VP2 and NS1 genes of CPV in feces from clinically CPV-infected dogs, and the nucleotide sequences were determined. The phylogenetic tree construction, homology and amino acid variation analysis of the deduced VP2 and NS1 amino acid sequences were performed using the software Meglign 7.0. Based on the deduced amino acid sequences analysis of VP2 gene, it showed that the epidemic strains CH-XY-7 and CH-XY-8 were New CPV-2a genotype. The epidemic strain CH-XY-5 was New CPV-2b genotype, and the epidemic strain CH-XY-1, CH-XY-2, CH-XY-3, CH-XY-4 and CH-XY-6 were CPV-2c genotype. There were 1（G299R） and 2（V84E and V153A） new variations in VP2 amino acids of epidemic strain CH-XY-3 and CH-XY-6, respectively. Based on the deduced amino acid sequences analysis of the NS1 gene, it showed that the epidemic strains CH-XY-5, CH-XY-6, CH-XY-7 and CH-XY-8 were divided. Into Group Ⅰ, with CH-XY-6 and CH-XY-8 each exhibiting a new amino acid variation（N356K in CH-XY-6, E583K in CH-XY-8）, CH-XY-1, CH-XY-2, CH-XY-3 and CH-XY-4 were divided into Group Ⅱ, and there are 5 new amino acid variants in epidemic strain CH-XY-3（N23S, F30L, N38D, R61G and Q307H）. It showed that the CPV prevalent in the southern Henan Province coexisted with multiple subtypes of CPV-2 and had mutated.