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The surge in gestational diabetes mellitus (GDM) globally requires a health system tailored approach towards prevention, detection and management. We estimated the prevalence of GDM using diverse recommended tests and diagnostic thresholds, and also assessed the risk factors and obstetric outcomes, including postpartum glycemia. Using a prospective cohort design, 446 singleton pregnant women without pre-existing diabetes did GDM tests in five hospitals in Ghana from 20–34 weeks using fasting plasma glucose (FPG), one-hour and 2-h oral glucose tolerance test (OGTT). Birth outcomes of 403 were assessed. GDM was diagnosed using six international diagnostic criteria. At 12 weeks postpartum, impaired fasting glucose (6.1–6.9 mmol/L) and diabetes (FPG ≥7.0 mmol/L) were measured for 100 women. Per FPG and 2-h OGTT cut-offs, GDM prevalence ranged between 8.3–23.8% and 4.4–14.3%, respectively. Risk factors included overweight (OR = 2.13, 95% CI: 1.13–4.03), previous miscarriage (OR = 4.01, 95% CI: 1.09–14.76) and high caloric intake (OR = 2.91, 95% CI: 1.05–8.07). Perineal tear (RR = 2.91, 95% CI: 1.08–5.57) and birth asphyxia (RR = 3.24, 95% CI: 1.01–10.45) were the associated perinatal outcomes. At 12 weeks postpartum, 15% had impaired fasting glucose, and 5% had diabetes. Tackling modifiable risk factors is crucial for prevention. Glycemic monitoring needs to be integral in postpartum and well-child reviews. [ABSTRACT FROM AUTHOR]