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Vitamin D deficiency is highly prevalent worldwide. 25-Hydroxy vitamin D (25(OH) vit D) is the major circulating source of vitamin D. Deficiency of vitamin D has been proposed as a new risk factor for cardiovascular disease, including stroke. We aimed to evaluate 25(OH) vit D (ng/mL) serum levels in patients with stroke according to severity and type in a cross-sectional study that out of 143 patients 78 were male and 65 were female. Serum concentration of 25(OH) vit D was measured at baseline by ELISA method. Stroke severity was assessed at admission using the NIHSS score. The mean of 25(OH) vit D serum level in male was 18.14 ng/mL and in female was 28.23 ng/mL. Data showed a significant difference between 25(OH) vit D serum level and sex (P = 0.009). At admission, 106 patients had ischemic stroke and 37 patients had hemorrhagic stroke with the mean of 22.52 and 23.31 ng/mL of 25(OH) vit D, respectively. Data presented no significant differences between 25(OH) vit D serum level and types of stroke. Furthermore, 40 patients (28%) had a minor stroke (0 ≤ NIHSS ≤ 5), 61 patients (42.7%) had a moderate stroke (6 ≤ NIHSS ≤ 14) and 42 patients (29.3%) had a severe stroke (15 ≤ NIHSS ≤ 42), with the mean of 21, 21.8, and 25.6 ng/mL of 25(OH) vit D, respectively. Data showed no significant difference between 25(OH) vit D serum level and severity (P = 0.61). This study showed that low 25(OH) vit D serum level is common in male but there was not any relation between serum level of 25(OH) vit D, type, and severity of stroke.