Background: Physical activity, assessed by accelerometers, has been proposed as a quantitative outcome measure for patients with DMD, but research is limited Objective: To assess the total amount and patterns of physical activity in patients with DMD using accelerometers. Methods: Physical activity was assessed in patients with DMD (n = 49, 13.6 ± 4.0-year-old) and age-and sex-matched healthy controls (n = 15, 14.0 ± 2.3-year-old) using wrist-and ankle-worn accelerometers. To assess the amount of activity, accelerometer recordings were converted into acceleration estimates (counts/min). Patterns of activity were assessed as the time that participants spent in sedentary, low-intensity, and moderate-to-vigorous physical activity categories. The sedentary category was divided into three (sedentary-1,-2, and-3) and the low-intensity into two (low-intensity-1, and-2) subcategories. Results: Physical activity across intensity categories differed between study groups (p < 0.001). Patients with DMD spent on average 98.8% of their daytime in the sedentary and low-intensity categories. Compared to non-ambulatory, ambulatory patients spent more time in sedentary-3 and low-intensity-2 subcategories (p < 0.001). Amount of activity was lower in all patients than controls (p < 0.05) and in non-ambulatory than ambulatory patients and controls (p < 0.001), but similar between ambulatory patients and controls. Activity measures in patients were significantly affected by age and ambulation status (p < 0.05) but not corticosteroid use. Conclusion: Patients with DMD spent most of their daytime in sedentary and low-intensity activities. Dividing these intensities into three and two subcategories, respectively, allows better characterization of activity patterns in DMD. Ambulation status and age but not corticosteroid use affected activity measures in patients with DMD.