Background: Canine parvoviral diarrhea is one of the most common viral infections in dogs. Canine parvoviral diarrhea is caused due to the virus, Canine parvovirus-2. Regular surveillance and knowledge of the emergence of new Canine parvovirus-2 viral variants in infected pet dogs is the prerequisite for the prevention and control of this disease. Methods: Deoxyribonucleic acids extracted from processed faecal samples of 50 diarrheic pet dogs were used to amplify Canine parvovirus-2 specific gene (Partial nucleotide region) by using a polymerase chain reaction test. Viral inoculum prepared from the positive faecal sample in the above test was adsorbed on Madin Darby Canine Kidney cell line of epithelial origin. The isolated virus was further characterized by using a molecular test: amplification refractory mutation system-polymerase chain reaction. Result: Deoxyribonucleic acid extracted from the processed faecal material of the dog tested positive for the presence of a Canine parvovirus 2-specific gene. The virus was successfully adapted to the Madin Darby Canine Kidney cell line and showed cytopathic effects on the very first passage. The isolated highly cytopathic virus Canine parvovirus-2 from the diarrheic pet dog was further characterized as Canine parvovirus-2a by using a molecular test.