Brucella canis, a facultative intracellular pathogen, is the causative agent of canine brucellosis. The diagnosis of canine brucellosis is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. In this study, recombinant B. canis Cu-Zn superoxide dismutase (SOD) was used as an antigen for the enzyme-linked immunosorbent assay (ELISA). The recombinant SOD showed a specific reaction with serum infected with B. canis in Western blotting and ELISA. These results suggest that ELISA using recombinant SOD is useful in screening for canine brucellosis.