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Abstract Background The purpose of the present study was to assess the validity and reliability of Form A of Multidimensional Health Locus of Control scales in Iran. Health locus of control is one of the most widely measured parameters of health belief for the planning of health education programs. Methods 496 university students participated in this study. The reliability coefficients were calculated in three different methods: test-retest, parallel forms and Cronbach alpha. In order to survey validity of the scale we used three methods including content validity, concurrent validity and construct validity. Results We established the content validity of the Persian translation by translating (and then back-translating) each item from the English version into the Persian version. The concurrent validity of the questionnaire, as measured by Levenson's IPC scale was .57 (P < .001), .49 (P < .01) and .53 (P < .001) for IPC, respectively. Exploratory principal components analysis supported a three-factor structure that items loading adequately on each factor. Moreover, the approximate orthogonal of the dimensions were obtained through correlation analyses. In addition, the reliability results were acceptable, too. Conclusion The results showed that the reliability and validity of Persian Form A of MHLC was acceptable and respectable and is suggested as an applicable criterion for similar studies in Iran.