We evaluated the potential effect of the lesion burden on the reproducibility of repeated lesion volume (LV) measurements from brain magnetic resonance imaging (MRI) scans of patients with multiple sclerosis (MS). Dual-echo, conventional spin echo brain MRI scans were obtained from 107 patients with MS. On proton density-weighted images, LV was assessed three times by the same raters, using a semi-automated, local thresholding technique for lesion segmentation. Mean LV (MLV) was 16.1 mL (range = 0.7-57.3 mL). The mean intra-observer coefficient of variation (COV) for the three measurement replicates was 2.6% (range = 0.2-7.2%). The intra-observer measurement variance (Var) increased with MLV and the fitted model was Var = 0.00187 MLV1.84. This indicates that LV measurements can be considered as measures whose variances are proportional to the square of their mean values, i.e., these measures have constant COV. Using a semi-automated, local thresholding segmentation technique, the reproducibility of LV measurements from brain MRI scans of patients with MS is not significantly influenced by varying lesion burdens.