Purpose: To determine the relationship between vaginal reflux (VR) and urinary tract infection (UTI). Methods: We performed a single center retrospective study of 191 girls, 36-months-old, with a diagnosis of febrile UTI, who underwent a voiding cystourethrography (VCUG) for assessment of vesicoureteral reflux (VUR) at our hospital. A control group of 51 girls, who underwent VCUG for assessment of congenital hydronephrosis or renal pelvis dilatation, without a UTI, formed the control group. The correlation between the presence and grade of VR and UTI was evaluated. Results: The prevalence rate of VR was higher in the UTI (42.9%) than control (13.7%) group (p0.05), with a higher VR severity grade in the UTI (mean, 0.64) than control (mean, 0.18) group (p0.05). On subanalysis, matching for age (UTI group, n=126, age, 5.28±2.13 months; control group, n=22, age, 4.79±2.40; p=0.33), the prevalence rate of VR remained higher in the UTI than the control group (43.65% versus 18.18%, p0.05), as did the VR grade (0.65 versus 0.22; p0.05). Presence and higher grade of VR were associated with UTI recurrence (p0.05). VR was correlated to urosepsis (p0.05). The renal defect rate among patient with VR(VR+/VUR+) was not different with those without VR (74% versus 52%; p=0.143) in VUR group. However, it was higher than VR+/VUR patients (74% versus 32%; p=0.001). If a child with VR+/VUR+ is exposed a UTI, the risk of renal defect increases. Conclusion: Occurrence of VR is associated with UTI recurrence, urosepsis and the risk of renal defect.