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Background: Many previous publications reported role of vitamin D insufficiency in pathogenesis of many disorders including psychiatric diseases such as autism and its mimic conditions. Few numbers of publications have studied role of vitamin D in development of autism in childhood. Aim: The aim this work was to study the status of vitamin D in autistic children by evaluation of serum levels of 25-hydroxy vitamin D (25 (OH) vitamin D) in Autistic pediatric patients and to study the beneficial role of vitamin D therapy as adjuvant therapy for this disease. Materials and methods: This study was conducted on forty pediatric patients who achieved the diagnostic criteria of autism depending on the 4th edition of Diagnostic and Statistical Manual of Mental Disorders (DSM-4). Thirty age and sex matched children served as control group. They were selected from pediatric and neuropsychiatric departments of Tanta Faculty of Medicine. All subjects were subjected to complete history taking, full medical examination, childhood autism rating scale (CARS) assessment and serum 25 (OH) vitamin D assay by enzyme-linked immunosorbent assay (ELIZA). Results: This study reported that 25 (OH) vitamin D levels in children with autism was significantly lower than the controls and it had significant negative correlation with CARS which was improved after vitamin D supplementation at a dose of 400 units daily for eight weeks. Conclusion: Vitamin D is insufficient in most of children with Autism, vitamin D supplementation can improve autistic features in these children.