Background: ASEAN job stressor questionnaire originally in English were determined the sources of job stress. Therefore, this study was conducted with the aim of measuring the validity and reliability of the Persian version of the ASEAN job stressor questionnaire (55 items) for physicians during 2008-2012. Methods: After translating the original version of the questionnaire from English to Persian using back-to-back translation method, its reliability and validity were examined. The study sample consisted of 780 doctors in hospitals in Tehran, Iran. The Spearman-Brown test and the correlation method were used to measure the validity and Cronbach’s alpha coefficient as internal consistency test were used to calculate the reliability. Results: Cronbach’s alpha coefficient was calculated to be 0.87. The reliability coefficient of the questionnaire was reported to be excellent by Spearman-Brown test and the correlation method of each question with the total score confirmed the reliability of the instrument. The results of factor analysis showed that there were two factors with a specific value greater than one that was able to explain more than 52.77% of the total score variance. Conclusion: The ASEAN job stressor questionnaire (55 items) is acceptable for determining the sources of job stress of physicians in the Iranian population.