There are few data focusing on the prevalence of vitamin D deficiency in tropical countries. We determined the vitamin D status in pregnant women and examined the factors associated with vitamin D deficiency. A cross-sectional study of 147 pregnant Thai women aged 18–45 years at Siriraj Hospital (a university hospital in Bangkok, Thailand) was undertaken. Clinical data and plasma levels of 25-hydroxyvitamin D [25(OH)D], intact parathyroid hormone (iPTH), calcium, albumin, phosphate and magnesium were obtained in pregnant women at delivery. The prevalence of hypovitaminosis D [defined as 25(OH)D <75 nmol/L] in pregnant women at delivery was 75.5% (95% confidence interval (CI), 67.7–82.2%). Of these, vitamin D insufficiency [defined as 25(OH)D 50–74.9 nmol/L] was found in 41.5% (95% CI, 33.4–49.9%) and vitamin D deficiency [25(OH)D <50 nmol/L] was found in 34.0% (95% CI, 26.4–42.3%) of women. The mean 25(OH)D concentration was 61.6±19.3 nmol/L. The correlation between 25(OH)D and iPTH was weak (r = –0.29, P<0.01). Factors associated with vitamin D deficiency by multiple logistic regression were: pre-pregnancy body mass index (BMI in kg/m2, odds ratio (OR), 0.88, 95% CI 0.80–0.97, P = 0.01) and season of blood collection (winter vs. rainy, OR, 2.62, 95% CI 1.18–5.85, P = 0.02). Vitamin D deficiency is common among pregnant Thai women. The prevalence of vitamin D deficiency increased in women who had a lower pre-pregnancy BMI and whose blood was collected in the winter. Vitamin D supplementation may need to be implemented as routine antenatal care.