Objective To investigate the early warning value of urinary selenium binding protein 1 (SBP1) in acute kidney injury (AKI) and its risk factor exposure, and compare it with urinary neutrophil gelatinase-associated lipocalin (NGAL). Methods Adult AKI inpatients and medical workers from the Department of Nephrology of the Second Affiliated Hospital of Xi’an Jiaotong University between April 2023 and April 2024 were selected. Patients who underwent percutaneous coronary intervention (PCI) in the Department of Cardiology of the Second Affiliated Hospital of Xi’an Jiaotong University were selected in June 2023. Patients who received cisplatin treatment in the Respiratory Department of the Second Affiliated Hospital of Xi’an Jiaotong University were selected in June 2023. Urinary SBP1 and NGAL levels of patients and medical workers were detected by enzyme-linked immunosorbent assay. Results A total of 14 medical workers and 36 AKI patients were included. Except for gender, alanine aminotransferase, aspartate aminotransferase, total cholesterol (P>0.05), there were statistically significant differences in other indicators between the medical workers and the AKI patients (P<0.05). The urine SBP1 [100.30 (71.50, 138.75) vs.75.60 (65.90, 80.08) pg/mL; U=2.918, P=0.004] and NGAL [423.70 (73.93, 839.80) vs. 14.80 (5.83, 29.98) ng/mL; U=4.668, P<0.001] levels in the AKI group were higher than those in the control group. But the area under the curve of receiver operative characteristic curve of urine SBP1 was smaller than that of urine NGAL (0.768 vs. 0.929). The urine SBP1 level in AKI patients was positively correlated with alanine aminotransferase, aspartate aminotransferase, serum creatinine, and serum glucose (P＜0.05), but negatively correlated with estimated glomerular filtration rate and total cholesterol (P＜0.05). A total of 14 patients who underwent PCI were included. The urinary SBP1/creatinine levels of PCI patients increased 6 hours after surgery compared to preoperative levels [(39.54 ± 8.00) vs. (19.34±2.90) pg/μmol; F=8.862, P=0.011]. The urea nitrogen level decreased 72 hours after surgery compared to preoperative levels (P=0.036), while there were no statistically significant differences in other indicators at other time points (P>0.05). There was no significant change in urinary NGAL levels before and after PCI treatment in patients. A total of 19 patients received cisplatin treatment were included. After cisplatin treatment, the level of urinary SBP1 increased compared to before treatment (P=0.024), while there was no significant change in the level of urinary NGAL after treatment compared to before treatment (P=0.350). After treatment, the levels of urea nitrogen (P=0.041) and cystatin C (P=0.002) increased compared to before surgery, while there was no statistically significant difference in blood creatinine and estimated glomerular filtration rate compared to before treatment (P>0.05). Conclusions Urinary SBP1 levels have certain diagnostic value for AKI, but the diagnostic efficacy is not as good as urinary NGAL. Urinary SBP1 is more sensitive to renal tubular injury caused by nephrotoxic drugs than urinary NGAL.