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Background: Ionizing radiation is one of the hazardous agents in the work place. It can cause serious and irreversible damage to the people exposed to it. Working in such environments can also causes chronic fatigue syndrome (CFS). The aim of this study was to investigate the prevalence of CFS in radiotherapists. Methods: This study selected all men and women exposed to X-ray as the exposed group and nurses working in the same hospital as the control group. The sample size was considered of 46 radiotherapists and 46 nurses. Data on demographic characteristics and chronic fatigue syndrome were collected by the questionnaire. Results: The mean CFS score in all participants was 10.64±4.77. About 17.39% of them had fatigue syndrome. There was no significant difference between two groups in terms of fatigue syndrome (p=0.47). radiotherapists with more than 20 image taking per day had higher mean CFS; however, this relationship was not statistically significant (P>0.05). Conclusion: Chronic fatigue is multifactorial phenomenon in health care personnel. Workload is more important factor in development of chronic fatigue in radiotherapists.