Objective: To determine repeatability of a wireless, inertial sensor–based lameness evaluation system in horses. Animals: 236 horses. Procedures: Horses were from 2 to 29 years of age and of various breeds and lameness disposition. All horses were instrumented with a wireless, inertial sensor-based motion analysis system on the head (accelerometer), pelvis (midline croup region [accelerometer]), and right forelimb (gyroscope) before evaluation in 2 consecutive trials, approximately 5 minutes apart, as the horse was trotted in a straight line. Signal-processing algorithms generated overall trial asymmetry measures for vertical head and pelvic movement and stride-by-stride differences in head and pelvic maximum and minimum positions between right and left sides of each stride. Repeatability was determined, and trial difference was determined for groups of horses with various numbers of strides for which data were collected per trial. Results: Inertial sensor–based measures of torso movement asymmetry were repeatable. Repeatability for measures of torso asymmetry for determination of hind limb lameness was slightly greater than that for forelimb lameness. Collecting large numbers of strides degraded stride-to-stride repeatability but did not degrade intertrial repeatability. Conclusions and Clinical Relevance: The inertial sensor system used to measure asymmetry of head and pelvic movement as an aid in the detection and evaluation of lameness in horses trotting in a straight line was sufficiently repeatable to investigate for clinical use.