OBJECTIVE Brucellosis is one of the most common zoonoses in the world, and occurs mainly in farmers, slaughterhouse workers, and veterinarians via direct or indirect contact with infected animals or their products. The clinical symptoms of human brucellosis are nonspecific, such as fever, headache, chills, and sweating. Diagnosis and treatment of brucellosis requires laboratory tests. Although the serum tube agglutination test (SAT) is the standardized gold method, it is laborious, time consuming, and requires a number of reagents. A microagglutination test (MAT) variant of the SAT or enzyme-linked immunosorbent assay (ELISA) is recommended for serological diagnoses. For the simple and rapid diagnosis of brucellosis, the MAT was standardized using samples for the SAT to define positive and negative categories, and we then compared the sensitivity and specificity of the MAT and ELISA. METHODS Thirty SAT-positive sera and 60 SAT-negative sera were used in this study. Antibody titers of ≥1:160 were considered positive readings in both the SAT and MAT. Brucella abortus antigens and Brucella-positive control antiserum were used in the SAT and MAT. ELISAs of IgM and IgG were performed according to the manufacturers' instructions. RESULTS The titers of the MAT differed according to antigen concentration. The optimal concentration of B abortus antigen was determined to compare the sensitivity and specificity between the MAT and SAT. The sensitivity and specificity of the MAT were 93.3% and 96.7%, respectively, for IgG with reference to ELISA, and 96.7% and 98.3%, respectively, for IgM. CONCLUSIONS The optimal concentration of antigen for the MAT was 1:10. The MAT is less time consuming and requires less antigen and serum than the SAT. The results of the MAT showed good agreement with those of ELISA. The results of this study suggest that the MAT could be useful for diagnosis of brucellosis.