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Ninety-three women with urodynamic stress incontinence (USI) and a mean age of 60.8 ± 10.7 (36–83) years were retrospectively enrolled. According to their VUDS, 31 (33%) were grouped into USI and detrusor overactivity (DO), 28 (30.1%) were grouped into USI and hypersensitive bladder (HSB), and 34 (36.6%) were controls (USI and stable bladder). The USI and DO group had significantly increased 8-isoprostane (mean, 33.3 vs. 10.8 pg/mL) and 8-hydroxy-2-deoxyguanosine (8-OHdG; mean, 28.9 vs. 17.4 ng/mL) and decreased interleukin (IL)-2 (mean, 0.433 vs. 0.638 pg/mL), vascular endothelial growth factor (mean, 5.51 vs. 8.99 pg/mL), and nerve growth factor (mean, 0.175 vs. 0.235 pg/mL) levels compared to controls. Oxidative stress biomarkers were moderately diagnostic of DO from controls, especially 8-isoprostane (area under the curve (AUC) > 0.7). Voided volume was highly diagnostic of DO from either controls or non-DO patients (AUC 0.750 and 0.915, respectively). The proposed prediction model with voided volume, 8-OHdG, and 8-isoprostane (cutoff values 384 mL, 35 ng/mL, and 37 pg/mL, respectively) had an accuracy of 81.7% (sensitivity, 67.7%; specificity, 88.7%; positive predictive value, 75.0%; negative predictive value, 84.6%). Combined with voided volume, urinary oxidative stress biomarkers have the potential to be used to identify urodynamic DO in patients with USI.