Brucellosis, a bacterial zoonoses caused by the genus Brucella is responsible for abortion and infertility in cow. Brucellosis is causing economic loss in dairy industries and prevalent worldwide including Bangladesh but limited studies are devoted to determine the prevalence and its association with reproductive factors of dairy cows in Bangladesh. Therefore, the present study was conducted to determine the seroprevalence of brucellosis in dairy cattle using screening test Rose Bengal test (RBT) and the positive sera were further confirmed by indirect- ELISA. For this purpose, a total of 400 serum samples from dairy cows with history of abortion and various reproductive disorders were collected from the Kurigram district of Bangladesh for the detection of Brucella antibody. The overall prevalence of brucellosis in dairy cattle was 2.25%. Brucellosis in cases of abortion and repeat breeding was 8.3% and 2.8%, respectively. The results shows higher prevalence of brucellosis in cases of abortion followed by repeat breeding, while there was no seropositive cases from other reproductive disorders. Age-wise sero-prevalence was found 3.0% in 2~3 years age group and 2.0% in 4~8 years age group. The prevalence of brucellosis in indigenous and cross-bred cattle was 3.6% and 1.7%, respectively. All the animals detected positive to brucellosis by RBT were not found to be positive by i-ELISA. However, the RBT might be a suitable screening test for the diagnosis of Brucella infection in field condition in Bangladesh. These data will help to develop effective disease prevention strategies.