This study was designed to estimate the contribution of repositioning inaccuracies to measured intracranial lesion volumes in multiple sclerosis (MS). Five patients with MS were each scanned 10 times, using spin-echo imaging (2,000/34/90), contiguous 5-mm slices, and different scanning positions. The maximum displacements from baseline were +/-4 degrees (AP rotation) and 3 mm (slice offset). Lesion volume was measured twice for each scan using a semiautomated contour technique. Measured lesion volumes ranged from 4,328 mm3 to 164,831 mm3. The mean intrarater coefficient of variation (CV) calculated for individual patients ranged from 1.1 to 4% (median, 1.7%). Using analysis of variance, the overall variance and CV due to altered scan position were greater than that due to rater error (repositioning CV 4.0%, intrarater CV 3.5%). The worst-case difference between volumes in the same patient ranged from 8.9 to 32% (median, 9.9%). Both rater and repositioning errors were greater for smaller lesion volumes. The maximum potential error due to repositioning inaccuracies is of a similar magnitude to the 5 to 10% expected change in lesion volume over 1 year. This study justifies continued careful attention to accuracy in repositioning for serial MR studies in patients with MS.