OBJECTIVE: To determine the accuracy of the 50-g, one-hour glucose screening test administered at 16 weeks of pregnancy for identifying women with gestational diabetes mellitus. STUDY DESIGN: Two hundred fifty-five women underwent 50-g, one-hour glucose screening tests at 16 weeks of pregnancy. Those with results > or = 135 mg/dL underwent 100-g, three-hour glucose tolerance tests. All patients without diagnoses of gestational diabetes during the second trimester of pregnancy underwent standard third-trimester glucose testing. RESULTS: Gestational diabetes mellitus was diagnosed in 25 patients. Glucose screening tests administered at 16 weeks of pregnancy identified 96% (24) of these patients. Patients with 16-week glucose screening test results > or = 135 mg/dL had a 55% risk of developing diabetes during pregnancy, while the risk was 0.6% for patients with 16-week test results < or = 110 mg/dL. Patients with 16-week glucose screening test results in the intermediate range, 111-134 mg/dL, had a 4.8% risk of developing diabetes during pregnancy. CONCLUSION: Glucose screening at 16 weeks of pregnancy is a useful alternative to third-trimester screening for gestational diabetes. The negative predictive value of screening test results < or = 110 mg/dL is 99.4%. The positive predictive value for screening test results > or = 135 mg/dL is 55%. This latter finding is superior to the 8.6-22% found during the third-trimester.