We investigated the seroprevalence and risk factors for Brucella seropositivity in cattle in Jordan. The sera from 671 cows were randomly collected from 62 herds. The antibodies against Brucella were detected using a Rose Bengal plate test and indirect ELISA. A structured questionnaire was used to collect information on the cattle herds' health and management. A multiple logistic regression model was constructed to identify the risk factors for Brucella seropositivity. The true prevalence of antibodies against Brucella in individual cows and cattle herds was 6.5% and 23%, respectively. The seroprevalence of brucellosis in cows older than 4 years of age was significantly higher than that in the younger cows. The seroprevalence of brucellosis in cows located in the Mafraq, Zarqa and Ma'an governorates was significantly higher than that of the other studied governorates. The multiple logistic regression model revealed that a larger herd size (odd ratio OR = 1.3; 95% CI: 1.1, 2.6) and mixed farming (OR = 2.0; 95% CI: 1.7, 3.7) were risk factors for cattle seropositivity to Brucella antigens. On the other hand, the use of disinfectants (OR = 1.9; 95% CI: 1.1, 2.1) and the presence of adequate veterinary services (OR = 1.6; 95% CI: 1.2, 3.2) were identified as protective factors.