Brucellosis is considered to be the most widespread zoonosis throughout the world. It has a serious implication on human health as well as on the economic development in a developing country like Bangladesh. The objective of the present study was to determine the seroprevalence and to delineate the risk factors for Brucella seropositivity in small ruminants in Mymensingh district of Bangladesh. In the present study, serum samples were collected from a total of 2456 small ruminants (1710 goat and 746 sheep) from 13 upazilla of Mymensingh district. The data related to age, sex and location were also collected using a questionnaire. Serum samples were screened using Rose Bengal Test (RBT) and Enzyme Linked Immunosorbent Assay (ELISA). Seroprevalence of brucellosis was 9.53% in goats and 9.92% in sheep on RBT test. In goat, the highest Brucella antibody was observed in Mymensingh sadar upazilla (13%) followed by Dhobaura upazilla (12.9%). On the other hand, highest ovine Brucella antibody observed in Haluaghat upazilla (13.04%) followed by Mymensingh sadar (12.5%). The prevalence was more in adults (55.2% in goats and 57% in sheep) than young (8.6% in goat and 8.1% in sheep) and more in female goats (41.1%) and sheep (39.2%) than male goats (14.1%) and sheep (18%). ELISA test showed 33.70% (31 out of 92 RBT positive samples) positive reaction of total RBT positive reactors. The result of this study can be useful to initiate and establish a program for controlling and prevention through test and slaughter, culling of infected animal from flock and vaccination.