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Several factors including ethnicity are known to influence 25(OH)D levels. The purpose of our study was to assess 25(OH)D levels among 1374 pediatric subjects of different ethnicity and to determine the prevalence of vitamin D deficiency and insufficiency among different ethnic groups. The prevalence of 25(OH)D ≤20 ng/ml was 44.2, 65.2, 69.2, 54.0, and 44.8 % among Caucasians, Africans, North Africans, Indians, and others, respectively ( P < 0.001). The median of 25(OH)D was 21.0 ng/ml (IQR = 14.0-29.6 ng/ml) for the cohort. Season of blood sampling, age, ethnicity, gestational age, birth weight, and z-score BMI were associated with 25(OH)D levels. Caucasians had higher median 25(OH)D levels than sub-Saharan Africans ( P < 0.001), North Africans ( P < 0.001), and Indians ( P < 0.001). There were no significant differences in the median 25(OH)D levels between ethnic groups among infants, whereas for children older than 1 year we found significant differences in 25(OH)D levels in the different ethnic groups, compared to Caucasians. Conclusion: Ethnicity was correlated with 25(OH)D levels among children older than 1 year. We found a high prevalence of vitamin D deficiency and insufficiency after the first year of life, and this was more remarkable in non-Caucasian children. [ABSTRACT FROM AUTHOR]