Gait impairments in persons with multiple sclerosis (pwMS) leading to decreased ambulation and reduced walking endurance remain poorly understood. our objective was to assess gait asymmetry (GA) and bilateral coordination of gait (BCG), among pwMS during the six-minute walk test (6MWT), and determine their association with disease severity. We recruited 92 pwMS (age: 46.6 ± 7.9; 83% females) with a range of clinical disability, who completed the 6MWT wearing gait analysis system. GA was assessed by comparing left and right swing times, and BcG was assessed by the phase coordination index (pci). Several functional and subjective gait assessments were performed. Results show that gait is more asymmetric and less coordinated as the disease progresses (p < 0.0001). Participants with mild MS showed significantly better BCG as reflected by lower PCI values in comparison to the other two MS severity groups (severe: p = 0.001, moderate: p = 0.02). GA and PCI also deteriorated significantly each minute during the 6MWT (p < 0.0001). GA and PCI (i.e., BCG) show weaker associations with clinical MS status than associations observed between functional and subjective gait assessments and MS status. Similar to other neurological cohorts, GA and pci may be important parameters to assess and target in interventions among pwMS. Multiple sclerosis (MS) is a degenerative, progressive, autoimmune disease of the central nervous system, often resulting in a continuous deterioration of walking 1. Hence, gait parameters, e.g. cadence, step-length, step-time, are impaired as compared to those measured in abled bodied individuals 1-4. This gait deterioration has been demonstrated as a decline in the ability to walk long distances, based on the 6-min or 500-m walk tests 5,6. One critical component of walking impairment is gait variability. Gait variability tends to change throughout the MS disease course, with greater variability in the higher levels of disability 7,8. Furthermore, gait variability is associated with increased fall risk 9. Human gait requires a high degree of symmetry and coordination. Gait asymmetry is associated with reduced walking velocity 10-12 and increased energy expenditure 13. Spinal cord injury is frequent in persons with multiple sclerosis (pwMS), noted in 83% by MRI and up to 99% at autopsy 14,15. Spinal cord injury is associated with lower extremity sensorimotor deficits and impaired ambulation. It was previously reported that walking velocity in pwMS was reduced when in vivo diffusion tensor imaging (DTI) of the cervical spinal cord reveals myelin and tissue injury within posterior columns (PC) and lateral corticospinal tracts (CST) 16. Since CST injury in pwMS is asymmetric 17 , we hypothesize that MS will be associated with increased gait asymmetry, since asymmetric lesions in the spinal white matter lesions have been shown to correspond to asymmetric motor function 18. Gait coordination is the ability to maintain a consistent phase-dependent cyclical relationship between different body segments or joints in...