Background: The Rapid Office Strain Assessment (ROSA) is one of the emerging posture assessment methods for the office workers. Limited studies have examined the validity of this method for the reliable assessment of the ergonomic risk factors among the office workers. We investigated the relationship between the ROSA and anthropometric dimensions, knowledge and behavior of the office workers in an Oil and Gas Company in Iran. Methods: This cross-sectional study was conducted in an oil company during 2016. All 259 office employees worked on the central headquarter and operational offices, completed the General Nordic, as well as the knowledge and behavior questionnaires. Then, the Rapid Office Strain Assessment (ROSA) and anthropometric dimension measurement were conducted for the participants. Data analysis was done for descriptive statistics, and Pearson correlation test using SPSS software (version 22). Results: The results showed an inverse association of the employees’ ergonomic knowledge and behavior with the prevalence of musculoskeletal disorders (p<0. 01). In addition, reduction of employees’ stress and duration of cellphone use, correcting the monitor height, and decreased ROSA scores, yielded decreased prevalence of musculoskeletal disorders (p<0. 01). Height, weight, age, gender, body mass index, work station, computer work, and job experience, had a significant correlation with the ROSA scores (p<0. 01). Conclusion: ROSA score along with other individual, workplace and work-related factors can be applied to assess the ergonomic risk factors in the office workplaces.