Objectives: (1) To examine the associations among ambulatory capacity, physical functioning and quality of life (QOL) in multiple sclerosis patients with different disability levels and compare the results to the healthy controls, and (2) to explain the relationship between QOL, disability status and disease duration. Methods: 112 multiple sclerosis patients and 50 healthy controls were selected for the study. Clinical and demographic data were recorded. Disability status (EDSS), walking ability and capacity (MSWS-12, T25FW), physical functioning (SF-36 /PF-10), QOL (MUSIQOL) and disease severity (MSIS-29) were evaluated. Outcome results of two subgroups with mild and moderate ambulatory impairment were analysed and compared with the control group. Results: There were a statistically significant difference between multiple sclerosis patients with low to moderate disability in MSWS-12, T25FW, SF-36 (PF-10) (p__ampersandsignlt;0.001), and MSIS-29 (p=0.018); additionally in the outcome results for EDDS (p__ampersandsignlt;0.001), MSWS-12 (p__ampersandsignlt;0.001), MSIS-29 (p=0.08) according to the physical functioning classification. Conclusions: The clinical impact of disability severity may contribute to impairments in walking, significantly influencing physical functioning in multiple sclerosis patients. QOL impairment may be emerged regardless of the disability level and disease duration.