Vitamin A deficiency [VAD] is the single most important cause of childhood blindness in developing countries. The present community-based study was done to assess the prevalence of VAD in Menia, Assiut and Sohag governorates of Upper Egypt and to analyze the nature of the problem to help in prioritization of the control intervention strategies. The study was conducted on preschool children aged 6-71 months. The two stage cluster sampling technique was used to choose sample of the study. The total sample size was 2700 children [30 clusters from each governorate urban and rural x 30 child from each cluster]. Data were collected from each child on socio-demographic characteristics, morbidity profile, dietary characteristics [breast feeding, qualitative and quantitative food consumption pattern], clinical eye signs of VAD and laboratory investigations [Hb estimation, serum retinol determination using HPLC and urine and stool analyses]. The prevalence of VAD was assessed using the biochemical indicator [serum retinol ≤ 0.7 μ mol/L]. Main findings revealed that VAD was mildly prevalent [4.8%] among preschool children of the study, being higher in Assiut [7.9%] followed by Sohag [4.4%] and Menia [2.4%], VAD was slightly more prevalent in urban sites and among males than females. Children from Assiut and Sohag, low intake of energy, protein and iron were all significantly associated with risk of VAD. An intermediate correlation existed between anemia and VAD. A set of recommendations was proposed summarized in vitamin A supplementation for preschool children and mothers, dietary diversification and increased consumption of vitamin A rich foods with improvement of other general public health measures.