Objective: To evaluate relative and absolute reliability and repeatability in assessing median nerve mobility at the level of the wrist and distal upper arm of the right upper extremity during wrist extension. Methods: Six healthy participants participated in the study. Median nerve mobility was captured three times at both sites using Sonocyte Turbo by two sonologists for a total of 72 video clips (36 for each site and 18 by each sonologist). Longitudinal movement was measured using Motion Tracking Analysis Program (MTAP) by the two assessors who were rehabilitation medicine residents. After one month, the assessors remeasured the longitudinal excursion of the median nerve of the previous video clips. Results: There was moderate agreement between the two sonologists of the median nerve mobility at the level of the distal upper arm and the wrist respectively. There was a moderate to almost perfect agreement between the two assessors’ readings in the mobility of the nerve at level of the distal upper arm and wrist for the first and second readings. Repeatability testing showed that there was variable agreement at the level of the distal upper arm and at the wrist. Conclusion: MTAP using fast template tracking with an adaptive template is a reliable tool that can be employed in the accurate assessment of median nerve mobility at the distal upper arm and wrist.