This study was performed to evaluate the seroprevalence of brucellosis among the risk group of population. A cross sectional study was carried out in the Department of Microbiology, Mymensingh Medical College in collaboration with the Department of Medicine under the Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, between the period from July 2007 to June 2008. A total of 210 samples were included in this study from risk group of population. A variety of serological tests have been applied for detection of antibodies against Brucella spp. Among the 210 subjects from risk group of population, 9(4.28%), 7(3.33%) and 7(3.33%) were positive for brucellosis by slide agglutination test, rose bengal plate agglutination test (RBPA) and standard tube agglutination test (STAT) respectively. Out of total specimen 10 positive and 29 negative serum as detected by slide agglutination, rose bengal plate agglutination test (RBPT) and STAT were also examined by Enzyme linked immunosorbent assay (ELISA) for detection of IgG. Among them, 10 were positive by ELISA IgG. Seroprevalence among occupational groups were 11.11% in veterinary personnel, 6.45% in dairy workers and 4.67% in animal farmers. This study indicated that brucellosis is not uncommon at rural area of Mymensingh region in Bangladesh. It was suggested to conduct community survey and to plan properly the measures of control and prevention.