Objectives: The aim of present study was to assess preliminary validity and reliability of the Health Related Quality of Life Questionnaire "KIDSCREEN-27" in a sample of Iranian students. Method: 551 students were selected from middle and high schools of Yazd city age 11 to 19, using multistage sampling. The questionnaire administered after pilot study and cognitive interview. Besides self report KIDSCREEN-27, 4^th version of Pediatric Quality of Life Inventory Generic Core Scales, General Health Questionnaire 28 item version (GHQ-28), Personal Well-being Index-School Children (PWI-SC), Multidimensional Students’ Life Satisfaction Scale (MSLSS), Cooper Smith Self-Esteem Inventory (CSSEI), Strengths and Difficulties Questionnaire (SDQ), and Socioeconomic Properties Questionnaire were performed. To examine validity of the questionnaire, construct validity (confirmatory factor analysis and discriminate validity) and convergent and divergent validity were employed, and using Cronbach’s alpha and test-retest (two week apart) coefficient its reliability was verified. Results: Floor effect was observed in psychological well-being, social support and peers, and autonomy and relation with parents, and ceiling effect was observed for all dimensions that were negligible. Confirmatory factor analysis revealed an appropriate fitness between the model and the data. There was moderate to strong correlation between KIDSCREEN dimensions, except for social support and peers, with similar dimensions in other questionnaires that assess the same constructs (p< 0.01). Significant differences in some KIDSCREEN dimentions were found between high and low socioeconomic, as well as among three and tree mental health (normal, borderline and abnormal) groups (p<0.05). Cronbach’s alpha coefficients for all dimensions, except for school environment dimension, were above 0.70 and test-retest reliability coefficients for all dimensions were strong (p<0.01). Conclusion: KIDSCREEN-27 has an appropriate validity and reliability in Iranian population.