Vitamin D deficiency is common in patients with polycystic ovarian syndrome (PCOS) and found to have multiple impacts on the disease process. Vitamin D status of women with or without PCOS in Bangladesh is largely unknown. This cross-sectional study was conducted in a tertiary level hospital of Bangladesh from January 2018 to April 2018 to address this lacuna. Sixty (60) newly diagnosed PCOS patients and 50 healthy controls aging ≥18 years were investigated for serum 25-hydroxy vitamin D [25(OH)D] level, fasting plasma glucose and fasting lipid profile in addition to their clinical and anthropometric profiles. None of the PCOS and the controls had sufficient 25(OH)D. Twenty five percent (25%) of PCOS patients were insufficient, 68.33% were deficient and 6.67% of were severely deficient of vitamin D; whereas in the control group the frequency was 12%, 50% and 38% respectively. PCOS patients had higher 25(OH)D than controls (17.53±4.6 vs. 13.79±6.1ng/mL, p<0.001). Although PCOS group had higher frequency of metabolic syndrome than control group (40% vs. 20%), 25(OH)D levels were similar in subjects with or without metabolic syndrome in both PCOS (16.82±4.74 vs. 17.99±4.49ng/mL, mean±SD, p=0.098) and control groups (14.06±5.94 vs. 13.73±6.20ng/mL, mean±SD, p=0.339). 25(OH)D level correlated with none of the clinical, anthropometric, metabolic and hormonal parameters in PCOS patients. Vitamin D deficiency is highly prevalent in Bangladeshi PCOS patients and healthy women of reproductive age.