Rabies is a zoonotic disease that is caused by rabies virus and transmitted only in mammals. Domestic dogs are the most common reservoir of the virus, which is associated with more than 99% human deaths caused by dog-mediated rabies in the world. Rabies is one of the most fatal diseases, but it is fully preventable in animals by vaccination. Serological test of rabies virus antibody for samples collected from dogs and cats in Seoul during 2017–2019 was carried out in this study. To investigate antibody seroprevalence of rabies virus for dogs and cats, 2,769 serum samples were taken from 2,408 dogs and 361 cats in various regions of Seoul during 2017– 2019. Antibodies to rabies virus were detected by an indirect ELISA. Of 2,769 tested animal sera, 934 (33.7%) were positive; 886 (36.8%) of 2408 dogs and 48 (13.3%) of 361 cats. Of 990 companion animals, 547 (55.3%) was positive and 387 (21.8%) of 1779 stray animals was positive. These results indicate that antibody seroprevalence to rabies virus is still not enough to prevent rabies and rabies vaccination is required to enhance the antibody seroprevalence for rabies. To improve the situation, much public awareness and policy is needed to prevent the rabies. In addition, reducing stray animals and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.