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The purpose of this study was to assess the concurrent validity and test–retest reliability of a sensor-based gait analysis system. Eleven healthy subjects and four Parkinson’s disease (PD) patients were asked to complete gait tasks whilst wearing two inertial measurement units at their feet. The extracted spatio-temporal parameters of 1166 strides were compared to those extracted from a reference camera-based motion capture system concerning concurrent validity. Test–retest reliability was assessed for five healthy subjects at three different days in a two week period. The two systems were highly correlated for all gait parameters ( r > 0.93 ). The bias for stride time was 0 ± 16 ms and for stride length was 1.4 ± 6.7 cm. No systematic range dependent errors were observed and no significant changes existed between healthy subjects and PD patients. Test-retest reliability was excellent for all parameters (intraclass correlation (ICC) > 0.81) except for gait velocity (ICC > 0.55). The sensor-based system was able to accurately capture spatio-temporal gait parameters as compared to the reference camera-based system for normal and impaired gait. The system’s high retest reliability renders the use in recurrent clinical measurements and in long-term applications feasible.