Background: Previous studies using ultrasound for diagnosis of carpal tunnel syndrome have reported on relatively small series of patients, leading to large standard deviations and/or confidence intervals for the mean cross-sectional area of the median nerve. The purpose of this study is to define the CSA of the median nerve in a large cohort of patients. Methods: Patients (n = 175) without history of carpal tunnel release were recruited. All participants were evaluated using the Carpal Tunnel Syndrome-6 questionnaire, a validated clinical diagnostic tool, with a score of 12 or greater considered positive for CTS. Ultrasound examination was performed on both wrists of all participants using a 13-6 MHz linear array transducer. Results: The mean median nerve CSA was significantly larger (P < .001) for patients with a positive (mean = 11.16, SD = 2.51) versus negative CTS-6 result (mean = 6.91, SD = 2.06). There was a significant correlation (.527, P < .001, n = 349) between CSA and CTS-6 score. Logistic regression analysis determined that a CSA of 10 mm2 optimized sensitivity and specificity at 80% and 88%, respectively. Accuracy was 87.9%. Conclusions: A significant difference in mean CSA was found between patients with and without CTS. Median nerve CSA showed a statistically significant positive correlation with CTS-6. Similar to prior studies, a CSA of 10 mm2 was determined to be the optimal cutoff. In this large series of patients, ultrasound was a sensitive, specific, and accurate test for confirmation of a clinical diagnosis of CTS.