Influenza A viruses (IAVs) are members of the family Orthomyxoviridae and genus Orthomyxovirus. Avian and mammalian species are the host of IAVs, which includes humans and dogs. Canine influenza virus (CIV) is an emerging pathogen that causes severe and acute respiratory diseases in dogs. This study monitored the antigen and antibody against CIV in dogs in the Republic of Korea (ROK) from 2016 to 2021. One thousand and seventy-two nasal swabs and 1,545 blood samples were collected from animal hospitals and animal shelters. Five nasal swabs in 2017 and seven in 2018 from stray dogs were positive for CIV according to RT-PCR. The prevalence of H3N2 CIV ranged from 9.5% to 24.8%, according to the hemagglutination inhibition (HI) assay. On the other hand, none of the serum samples from 2018 to 2021 showed seropositivity against the avian H5, H7, and H9 viruses. The HI titers for H3N2 ranged from 16 to 512. The distribution of HI titer 16–32 was 57.6% in seropositive samples. The pet dogs were vaccinated against CIV, but the stray and military dogs were unvaccinated. In 2017 and 2018, the seroprevalence of CIV in stray dogs was higher than in the other years, and viral RNA was detected in nasal swabs. It may mean previous exposure of stray dogs to CIV. With the increasing number of pet dogs and the close contact between humans and dogs, canines could serve as an intermediate host for transmitting IAVs to humans. Therefore, continuous surveillance of CIV is needed for public health and the potential emergence of novel zoonotic viruses.