Objective: Patellofemoral pain syndrome (PFPS) is a condition that is characterized by patellar discomfort or pain that is aggravated during certain activities such as ascending/descending stairs. The Patellofemoral Disability Index (PDI) was developed to assess the effect of pain on functional activities in individuals with PFPS. The objectives of the current study were to determine the internal consistency, test-retest reliability, and validity of this index. Design: Cross-sectional study. Methods: Forty-one subjects who had PFPS with a mean age of 28.8±5.0 years and a mean body mass index of 25.6±4.7 kg/m² participated in the study. All subjects were concurrently enrolled in a clinical trial for which they were instructed to complete hamstring-resistance exercises for 4 weeks. Over the course of the intervention, they completed both the PDI and the Oswestry Disability Index (ODI) at baseline after two weeks, and after four weeks. Pearson correlation coefficient was used to assess the criterion validity. Cronbach’s α was used to examine the internal consistency. Intraclass correlation coefficients with 95% confidence interval were computed to examine test-retest reliability. Results: Subjects’ responses within both the PDI and the ODI yielded Pearson correlation coefficient values that were positive and highly significant (range, 0.73-0.97; p<0.001). There was a high level of internal consistency (Cronbach’s α≥0.8), with the exception of stair climbing (Cronbach’s α=0.65). Intraclass correlation ranged from 0.87 to 0.92, indicating high levels of test-retest reliability. Conclusions: The PDI is a valid, reliable, and feasible method of assessing pain and functional ability in patients with PFPS.