Objective: To evaluate the clinical potential of urine prostatic exosomal protein (PSEP) as a diagnostic biomarker of chronic prostatitis (CP). Materials andmethods: Using an enzyme-linked immunosorbent assay kit, urine PSEP levels were detected in 103 control cases as well as 283 cases of CP, with 82 cases fulfilling the definition of the USA National Institutes of Health category II (NIH-II), 108 cases of NIH-IIIa and 93 cases of NIH-IIIb. The values of age, body mass index, prostate volume, serum prostatic specific antigen (PSA) urine PSEP levels, and seminal parameters were analyzed. Results: The PSEP levels were significantly higher in patients of NIH-II (2.09 [2.35] ng/mL), NIH-IIIa (1.80 [2.95] ng/mL) and NIH-IIIb (1.64 [2.48] ng/mL) compared to the value of 0.24 (0.76) ng/mL in the controls. ROC identified a cutoff value of 1.387 ng/mL, with a sensitivity of 59.0% and specificity of 94.2%. The area under the ROC curve was 0.833. PSEP levels positively correlated with serum PSA levels in the NIH-IIIb group, and with EPS WBC count in the NIH-IIIa group, and with semen WBC count in each CP subgroups but negatively correlated with sperm motility in both the NIH-IIIa group and the NIH-IIIb group. Conclusion: Urine PSEP could be a potential biomarker for CP.