The musculoskeletal problems (MSP) vary among different occupations since they had different characteristics and physical workloads. Therefore, it is important to know the difference between the occupational groups to design preventing physiotherapy interventions. To investigate the prevalence differences of MSPs and related physical workload among hospital staff. In this cross-sectional study, 416 hospital staff completed the Nordic Musculoskeletal Questionnaire for MSP and Physical Workload Questionnaire for assessing the physical workload. One-year prevalence of low back, neck, upper back, and shoulders were 73.8%, 59.9%, 59.4%, and 52.2%, respectively. The most preventing MSPs from work found in the low back (39.2%), upper back (26.7%), and the neck (24.5%). MSP of low back impacted nurses the most with a 1-year prevalence of 81.3% and 57.1% of nurses were prevented from working. Nurses, service and cleaning staff had significantly more physical workload than secretaries and physicians. MSP of low back had the highest prevalence among hospital staff and it was the leading cause which prevented from working. Nurses were the most in danger in terms of MSPs among hospital staff. Physical workload was significantly higher in nurses, service and cleaning staff than secretaries and physicians.