Background: Gestational diabetes mellitus (GDM) represents the most common metabolic complication during pregnancy. GDM is associated with maternal and fetal complications. Approximately 7% of all pregnancies are affected by GDM, resulting in more than 200,000 cases worldwide annually, and the prevalence may vary from 1% to 14% among all pregnancies. Accordingly, this study attempted to determine the prevalence and some risk factors of GDM. Materials and Methods: This hospital-based cross-sectional study was carried out at Boo-Ali hospital in Tehran, the capital of Iran. Four hundred non-diabetics pregnant women with a gestational age of 24-28 weeks who attended the Boo-Ali hospital outpatient department were included in our study. Demographic and anthropometric data including age, gravida, para, gestational age, family history of diabetes, history of GDM, weight, height, and body mass index (BMI) were collected. Blood samples were collected from the women at 24-28 weeks to diagnose GDM by oral glucose tolerance test (OGTT). We measured the 25-OH-D level in participants at 24-28 weeks. Results: Among the 400 pregnant women, 46 (11.5%) had GDM based on OGTT, and the mean age of GDM women were 30.78± 5.96 years. Among selected variables, BMI ≥25kg/m2, family history, and GDM history were associated with increased risks of GDM (odds ratio=2.49, 95% confidence interval [CI] 1.22–5.07;3.52, 95% CI 1.84–6.70; 19.57, and 95% CI 6.16–62.17, respectively). The association was more robust in the positive GDM history of women. Conclusion: High prevalence of GDM highlights more attention of health-care givers in screening pregnant women with risk factors. BMI as a modifiable risk factor for GDM needs more attention, and positive family history and previous GDM history should be considered in the core activities of pregnant women. [GMJ.2020;9:e1642] DOI:10.31661/gmj.v9i0.1642