Canine parvovirus, caused by a haemagglutinating CPV, is one of the most important acute viral infectious diseases of pups, had been prevalent in the country. In the present study, 50 faecal samples, from clinically suspected cases of parvovirus diseases dogs were collected from pet center of UVAS, Lahore. All the samples collected from the infected dogs were analyzed for complete blood count. There were decrease in the value of certain blood parameters including TEC, TLC, PCV. ESR ,Hb, Plt, MCV,MCH,MCHC. The samples were diluted and centrifuged to collect the supernatant. Being a haemagglutinating (HA) virus, the pre-filtered supernatant from all suspected samples was checked for HA activity using 1% washed chicken erythrocytes. Out of 50 samples, 35 samples were found HA positive. All the HA positive and negative samples were processed for extraction of viral DNA with Genomic DNA purification kit. The net isolated DNA samples were subjected to Polymerase Chain Reaction (PCR) by using specific primers (developed against the variable genomic region of the capsid protein of the DNA) of the CPV. One of the selected primer pair amplified the genomic region present in the CPV-2b. Optimization of PCR was done for the molecular level daignosis of canine parvovirus and this technique was previously not available or standardized in local conditions of Pakistan. With the use of primer pair, best results were found on following optimizing conditions like annealing temperature 55 0C, primers concentration (reverse and forward) 30 pmoles, Magnesium Chloride concentration 2.5mM, DNA (Template) volume 5 µl, Taq DNA polymerase (12.5U) and 200 µM of each dNTPs. The PCR product was analyzed for the banding pattern of 681 bp amplified by the primer pair against the standard DNA ladder (100bp) by using horizental agarose gel electrophoresis. Four of the HA negative samples were amplified by the PCR reaction. and the presence of CPV 2a and CPV 2b in Pakistan as the prevalent pathogenic strains. This study has been a first step for the molecular diagnosis of canine parvovirus in local conditions of Pakistan. It is hoped that this study will pave the way for further advanced studies on this topic.