Musculoskeletal disorder rates among healthcare workers are high compared to other occupational groups. Studies indicate a higher prevalence of musculoskeletal pain in women as compared to men in most productive sectors. The objectives of our study were to assess the prevalence of upper-limb musculoskeletal pain in male and female employees of the Belo Horizonte municipal Health Department, and to identify associated factors, considering individual and occupational characteristics, by gender.