To understand the current situation of canine parvovirus (CPV) epidemic strains in Jilin Province and to analyze the molecular biological characteristics and genetic evolution of the epidemic strains, 44 fecal or intestinal tissue samples detected by canine parvovirus test strips in Changchun and Liaoyuan City, Jilin Province, from February 2018 to November 2019 were collected for cloning, sequencing and genetic evolution analysis of the gene encoding the virus structural protein VP2. The results showed that 44 of the 44 samples were CPV-2 positive by PCR, among which CPV-2c was the dominant variant (70.4%). In addition, new-CPV-2a (18.2%), new-CPV-2b (9.1%) and CPV-2 (2.3%) also existed. This is the highest frequency of CPV-2c subtypes observed in canine populations in Jilin Province so far. In addition to the substitutions of Ala5Gly, Phe267Tyr, Tyr324Ile, and Gln370Arg in the VP2 protein, a novel variant with an Arg481Lys mutation was observed in a CPV-2c strain. Hence, there is a subsequent need for further and extensive epidemiological investigation and eventual adaptation of current vaccines.