The role of physical therapy in health promotion in workplace is increasingly important. Musculoskeletal pain is a common problem for hospital personnel that physical therapists may assist in its prevention and management. However, the information concerning risk factors related to musculoskeletal pain in hospital personnel has been limited. This study therefore aimed to examine the prevalence of musculoskeletal pain in hospital personnel and to explore its risk factors. The information will help provide the reference for planning health promotion program for hospital personnel. This study prospectively enrolled employees from a hospital in northern Taiwan. All subjects were examined their musculoskeletal pain using the Nordic Musculoskeletal Questionnaire and the physical activity using the International Physical Activity Questionnaire- Taiwan version. Their fitness including waist girth, muscle strength and endurance, flexibility, balance and cardiopulmonary index were assessed by physical therapists. Logistic regression analysis was used to examine the associations of potential influencing factors (i.e., sex, occupation, physical activity and fitness) with the occurrence of musculoskeletal pain. This study included 645 hospital employees, 80% being females with mean age of 34 years and 20% being males with mean age of 36.4 years. 464 employees (72%) reporting musculoskeletal pain in the past one year, 55% encountered upper quarter pain and 43% encountered lower quarter pain. 443 employee (69%) reported that musculoskeletal pain influenced work. Multivariable logistic regression analysis revealed that individuals having poor muscle endurance were more likely to have pain over the lower quarter (odds ratio [OR]=1.8, p=0.03). Females had higher risk of sustaining upper quarter pain than males (OR=2.2, p=0.001). Furthermore, employees with prolonged sitting time during work in the past 7 days were more likely to have upper quarter pain (OR=1.001, p=0.035). Our results indicated association of poor abdominal muscle endurance with lower quarter pain; and relation of female and prolonged sitting during work with upper quarter pain. Through participation in the health promotion committee and activities, physical therapists can provide evidence-based services to help implement the policy of health promotion.