Fluorodeoxyglucose (FDG) positron emission tomography (FDG PET/CT) can be used to identify and localize infection in patients with vascular graft infections (VGI). We aimed to evaluate the diagnostic accuracy of 18F-FDG PET/CT by defining thresholds for standardized uptake value (SUV) and tissue-to-background ratio (TBR) that would accurately identify the presence of vascular graft infection. Patients with suspected VGI were prospectively recruited and underwent 18F-FDG PET/CT scans. Diagnosis was based on clinical, laboratory and radiologic findings, and blinded to the results of the PET/CT scan. Receiver operator characteristics (ROC) curve analysis was done to determine optimal thresholds for SUV and TBR. Our final cohort consisted of 28 patients with suspected VGI (mean±SD age 67±10years, 61% men), of which 15 patients (54%) had definitive VGI. The cohort included 61% prosthetics grafts and 39% stent-grafts. The type of graft included in this study were biologic (4%), Dacron (64%) and Polytetrafluoroethylene (32%). The location of the implanted grafts was aortic (54%) and peripheral arterial reconstruction (46%). The location of the peripheral graft was 77% in lower extremity and 23% in the upper extremity (arterio-venous grafts for dialysis access). Using ROC analysis, SUV max of 4.5, SUV mean of 3.7, and a TBR of 1.6 gave the best balance between sensitivity and specificity (93%/92%, 100%/92% and 93%/92%, respectively). All thresholds had an area under the curve ≥0.93 and correct reclassification rate ≥93%. Our data suggests that FDG PET/CT can be used to reliably and accurately diagnose VGI. The dual anatomic-physiologic information from FDG PET/CT can complement clinical diagnosis particularly in uncertain cases.