This study evaluates the effects of a rehabilitation program based on reflex locomotion therapy (RLT) on balance, gait, and fatigue in patients with multiple sclerosis (MS). Twenty-three patients diagnosed with MS participated in this study. Reversal design was carried out. The assessment tools included the Berg Balance Scale (BBS), the Performance Oriented Mobility Assessment (POMA), the Fatigue Severity Scale (FSS) and the instrumental analysis of the gait recorded by Vicon Motion System®. We analyzed spatio-temporal parameters and kinematic variables of the hip, knee, and ankle joints. Additionally, the Client Satisfaction Questionnaire (CSQ-8) was administrated. We did find a significant improvement in balance and gait tools after the RLT period. Regarding instrumental analysis, the statistical analysis of spatio-temporal parameters showed a significant improvement in stride length, double support, and velocity after the RLT period. Concerning kinematic parameters, the analysis showed improvements in hip and knee range of motion (ROM) after RLT period. RLT could improve gait and balance in patients with MS. The patients reported a high level of satisfaction with the therapy received.