|Objective To determine the highest 50-g glucose challenge test (GCT) value that indicates no further diagnostic test is needed to confirm a diagnosis of gestational diabetes mellitus (GDM) under the criteria of National Diabetes Data Group (NDDG) or the Carpenter and Coustan (C&C) and fasting glucose thresholds from the International Association of Diabetes and Pregnancy Study Group (IADPSG). Methods We collected the 50-g GCT results from 16,560 pregnancies and identified 2,457 gravidas with positive 50-g GCT (≥130 mg/ dL) values who underwent the 100-g glucose tolerance test. We investigated GDM prevalence in pregnancies with positive 50-g GCT according to the respective diagnostic thresholds and determined the 50-g GCT cutoff values with 100% positive predictive value for GDM under each diagnostic threshold. Results Twelve point five percent (306/2,457), 20.0% (492/2,457), and 9.6% (235/2,457) met the diagnostic criteria of GDM with the application of NDDG, C&C criteria, and fasting glucose thresholds from IADPSG (≥92 mg/dL), respectively. We also found that the prevalence of GDM increased with increasing 50-g GCT values using each diagnostic criterion. Importantly, we identified that all subjects with a 50-g GCT value ≥223, ≥217, or ≥228 mg/dL can be exclusively diagnosed as having gestational diabetes according to the criteria of NDDG, C&C, and fasting glucose thresholds from IADPSG, respectively. Conclusion We propose that women with a 50-g GCT screening value ≥228 mg/dL can be reliably omitted from further confirmative tests for GDM, such as 100- or 75-g glucose tolerance test.