BACKGROUND AND OBJECTIVES: Timed 25-foot walk (T25FW) is a common and reliable measure of ambulatory impairment in people with multiple sclerosis (MS). There is a lack of evidence evaluating the treatment effects on T25FW in MS. This study aimed to estimate treatment effects on T25FW in MS through a systematic review and meta-analysis. METHODS: From PubMed, Embase, and Cochrane Library databases, 41 articles were eligible for this study, including 21 parallel-arm and 20 single-arm studies. A multilevel meta-analysis summarized the comprehensive treatment effects on T25FW with odds ratios (OR) and standardized mean changes (SMC) for parallel- and single-arm studies, respectively. RESULTS: In the meta-analysis, the overall comprehensive treatment effect on T25FW was confirmed, showing a concurrent effect (OR=1.28
95 % CI=1.10-1.48, P=0.0038) in parallel-arm studies and a longitudinal effect (SMC=0.20
95 % CI=0.03-0.36, P=0.023) in single-arm studies. Specifically, ocrelizumab demonstrated the most concurrent effect (OR=1.60
95 % CI=1.17-2.21, P=0.0036), though based on a single study with binary T25FW scores. On the other hand, fampridine had the most longitudinal effect (SMC=0.24
95 % CI=0.12-0.36, P=0.0022), based on ten studies. CONCLUSION: In the current multilevel meta-analysis, we provided comprehensive treatment effects for T25FW in MS, confirming the better effects of ocrelizumab and fampridine.