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Aim: To find an optimal threshold level with higher sensitivity and specificity for screening of gestational diabetes mellitus (GDM) in Turkish pregnant women. Materials and methods: This was a retrospective study. Screening for GDM was performed in all pregnant women between 24 and 28 weeks of gestation using the 1 h 50 g glucose challenge test (GCT) with a subsequent 3 h 100 g oral glucose tolerance test (OGTT) for confirmation if screened positive. The glucose values obtained were analyzed by both the Carpenter and Coustan (C&C criteria) and National Diabetes Data Group (NDDG) criteria. Results: There were 808 women meeting the study inclusion criteria. There were 66 (8.1%) women diagnosed with GDM using the C&C criteria and 45 (5.7%) using the NDDG criteria. The best cut-off point for GCT was 132 mg/dL for detecting GDM. No diabetes was found below the glucose level of 130 mg/dL. Conclusion: GCT is suitable for screening of Turkish women, but place of residence as well as race must be taken into consideration to establish the best cut-off level of GCT, since ethnic and environmental factors may contribute to the occurrence of GDM. [ABSTRACT FROM AUTHOR]