Uterine fibroids are monoclonal tumors of uterine smooth muscle cells. They are found in 20-40% of females in the reproductive age group. Ovarian hormones play an important role in its etiology. Recent studies have shown that low serum 25(OH)vitamin D levels are associated with increased risk of uterine fibroids. To investigate the association between serum vitamin D levels and uterine fibroids.A case control study was conducted in a tertiary care hospital among 183 women between the age of 18-50 years with uterine fibroids who attended obstetrics and gynecology department and 183 women of the same age group who had no uterine fibroids attending comprehensive health checkup clinic. Serum 25(OH)vitamin D levels were estimated in both groups by electrochemiluminescence immunoassay.Statistical analysis was performed using IBM SPSS version 20.0. P value of <0.05 was considered statistically significant. The mean serum 25(OH) vitamin D levels of women with uterine fibroids were 15.03 ± 6.996 ng/mL and 16.629 ± 8.020 ng/mL in women without uterine fibroids. There was no statistically significant difference in serum 25(OH) vitamin D levels between women with and without uterine fibroids (P-0.132).Serum 25(OH) vitamin D levels were found to be deficient in both groups irrespective of the presence or absence of uterine fibroids. Hence, it can be concluded that the study conducted in 366 women could not find a direct association between low serum 25(OH) vitamin D levels and uterine fibroids.