Four strains of hybridomas that could stably secrete monoclonal antibodies against canine distemper virus（CDV）and canine parvovirus（CPV）were screened by cell fusion and indirect ELISA，based on which，a colloidal gold-based dual detection card that could simultaneously detect CDV and CPV was developed，then its specificity，sensitivity and accuracy were tested. The results showed that the four strains of hybridomas could be stably passaged，the purity of monoclonal antibodies secreted was high，and all the titers were above 1:320；the prepared card，with strong specificity，could produce specific strips with CDV and CPV only，but failed with other canine viruses；high sensitivity was observed，and the minimum detection limit could reach 102 copies/μL；and good accuracy made the coincidence rate with the results by fluorescent quantitative PCR higher than 88.0%，and that with the single virus detection card higher than 97.2%. In conclusion，the prepared card was with high sensitivity，specificity and accuracy as well as other advantages such as simple operation，visual interpretation，easy preservation of results，and working without any special instruments and equipment，which could be used for rapid clinical diagnosis and massive detection of the two diseases.