Developmental coordination disorder (DCD) is a condition that results in an impairment of gross and/or fine motor coordination. Compromised motor coordination contributes to lower levels of physical activity, which is associated with elevated body fat. The impact of elevated body fat on motor coordination diagnostic assessments in children with DCD has not been established. The purpose of this study was to determine if relative body fat influences performance on the Movement Assessment Battery for Children, 2nd Edition (MABC-2) test items in children with and without DCD. A nested case–control, design was conducted within the Physical Health Activity Study Team longitudinal cohort study. The MABC-2 was used to assess motor coordination to categorize cases and matched controls. Relative body fat was assessed using whole body air displacement plethysmography. Relative body fat was negatively associated with the MABC-2 “balance” subcategory after adjusting for physical activity and DCD status. Relative body fat did not influence the subcategories of “manual dexterity” or “aiming and catching”. Item analysis of the three balance tasks indicated that relative body fat significantly influences both “2-board balance” and “zig-zag hopping”, but not “walking heel-toe backwards”. Children with higher levels of relative body fat do not perform as well on the MABC-2, regardless of whether the have DCD or not. Dynamic balance test items are most negatively influenced by body fat. Health practitioners and researchers should be aware that body fat can influence results when interpreting MABC-2 test scores.