Vitamin D deficiency (VDD) is one of the serious and highly debatable public health problems affecting at least one billion of world population. This study objected to evaluate Vitamin D status in adult population of both sexes residing in different geographical areas of Kazakhstan and to elucidate the possible contributing factors related to VDD. This cross-sectional study covered 6 regions of Kazakhstan and applied the systematic random sampling to recruit 1347 healthy adults (of whom 819 were females) with mean age 44 ± 14 years. The concentration of 25-hydroxy vitamin D (25-OHD) was measured from May 2018 to August 2018 with Architect 25OH Vitamin D assay (Abbott Ireland Diagnostics Division Lisnamuck, Longford Co. Longford Ireland). Vitamin D deficiency was defined as 25-OHD values not exceeding 20 ng/mL as a reference threshold in healthy population. The median serum 25(OH)D concentrations in all studied regions of Kazakhstan were below the reference threshold (20 ng/mL). The lowest range of vitamin D (<10 ng/mL) was observed more commonly in females (34.6 % - 283) as compared to males (16.7 % - 88) and was significantly higher in Asians (33.2 % - 352) in contrast with Caucasians (6.7 % - 19) (χ2 = 177,939; D.f. = 3; p-value=<0,001). The proportion of severe VDD was higher in individuals with low body mass index (31.1 % - 188) vs. individuals with high body mass index (18.7 % - 50). In this study individuals aged 60 years and older had the most favorable situation with 25-OHD concentrations since these were normal in 14.4 % of observations (χ2 = 26,589; D.f. = 6; p-value=<0001). Studying the prevalence of VDD is an important public health task. Further research is needed to understand the epidemiology of VDD in more details to tailor intervention programs.