Canine parvovirus-2 (CPV-2) has been the most prevalent causative agent of enteritis in puppies throughout Egypt and all around the globe. Following its first identification in 1978, CPV-2 became swiftly supplanted with three antigen subtypes: CPV-2a, CPV-2b, and CPV-2c. The three subtypes of CPV-2 have been reported in Egypt. Therefore, this study aimed to monitor clinical findings, laboratory changes in puppies suffered from CPV, and its molecular characterization using polymerase chain reaction (PCR) and DNA sequencing to further understand the subtypes of CPV-2 that are present in dogs in Ismailia, Egypt. The clinical signs of infected puppies were vomiting, dehydration, and foul-smelling bloody diarrhea. Microcytic hypochromic anemia and thrombocytopenia were evident in association with leucopenia, neutropenia, and lymphopenia. In addition, hyponatremia, hypokalemia, hypochloremia, hypocalcemia, and hypophosphatemia were also recorded in diseased puppies. All 47 tested fecal samples were positive to CPV-2 when examined by PCR and gel electrophoresis. The sequence analysis of CPV-2 showed that the most common subtype is CPV-2a that is followed by CPV-2b as well as CPV-2c with percentages of 71.4%, 14.3%, and 14.3%, respectively. In conclusion, CPV-2a was the predominant subtype isolated from the examined puppies and was responsible for the characteristic clinical signs and laboratory changes in diseased dogs. Moreover, epidemiological investigations are needed to determine the prevalence of CPV serotypes in Egypt.