Background: Canine parvovirus (CPV) emerged in 1970s as a highly infectious disease. CPV modifi ed live vaccines have been widely used to control the disease. It is urgent to develop specifi c monoclonal antibodies to differentiate fi eld virus from vaccine virus in vaccinated dogs. Methods: In this study, female BALB/C mice were immunized with a commercial CPV vaccine strain. Two monoclonal antibodies (MAbs) 10H4 and 10B11 were made by hybridoma technique, and screened by hemagglutination inhibition (HI) assay. Results: MAb 10H4 reacted with both CPV fi eld and vaccine strains, and MAb 10B11 only recognized fi eld but not vaccine strains by the results of HI, indirect immunofl uorescence (IFA), and virus neutralization tests. Conclusion: Therefore, these two MAbs may work as useful tools to study the CPV pathogenic mechanisms and to develop diagnostic reagents.