Musculoskeletal pain is one of the reasons for a musician's inability to play an instrument. An assessment of the localization, intensity, and interference of those complaints is important among musicians because of the risk of occupational disease. Assessment by validated tools is especially important and serves as an indicator to take up proper preventive or treatment actions. The aim of the study was to assess the prevalence of playing-related musculoskeletal disorders (PRMDs) and the relationships with PRMDs' impact on playing musical instruments among Polish professional orchestra musicians. The study was conducted on a group of professional orchestra musicians (age 37.19 ± 9.62 years), 99 (55%) women and 83 (45%) men. Work experience in professional orchestra was 18.3 ± 9.83 years and the reported years of playing musical instrument were 31.4 ± 9.50. The study used the online version of a musician-dedicated questionnaire, Musculoskeletal Pain Intensity and Interference Questionnaire for Musicians of the Polish Population (MPIIQM-P). In the study group, 89.9% of women and 83.1% of men had experienced playing-related pain in their lifetime. The most intense pain among the group with current playing-related musculoskeletal complaints ( = 85) was located in the neck area for 19 (22.4%). The results of the study indicate a high prevalence of playing-related musculoskeletal problems among professional musicians.