Canine adenovirus (CAV) circulates as two distinct serotypes, CAV-1 and CAV-2, which are antigenically related but differ in their clinical manifestations. CAV is one of the important viral agents in the etiology of canine gastroenteritis. Here, we report the molecular surveillance and genetic characterization of CAV from clinical cases of canine gastroenteritis. A total of 302 fecal/rectal swabs were collected from dogs presented with gastroenteritis at various clinics in and around Hyderabad, India during 2018–19. These samples were tested for CAV using polymerase chain reaction with primers designed for the CAV E3 gene and the virus was isolated from positive samples. CAV-2 nucleic acid was present in 4.9% of the test samples. The partial sequence analyses of the E3 gene of the CAV-2 isolates revealed a frameshift mutation by insertion of nucleotide “G” at 1077 position of E3 gene, which resulted in an extension of the polypeptide chain by eleven amino acids. As a result, isolates from the current study formed a novel group, and the virus that was previously subdivided into two groups worldwide is now categorized under three. The study identifies a novel group of CAV-2 circulating in India providing an updated information regarding CAV-2.

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