Professional rickshaw driving is a seemingly sedentary occupation and involves many risk factors for work-related musculoskeletal disorders (WMSD). To assess the prevalence of work-related musculoskeletal disorders and its associated risk factors among professional rickshaw drivers. 263 rickshaw drivers were voluntarily recruited from Aligarh, Uttar Pradesh, India. MSD symptoms in the past 12 months and last 7 days were assessed using a self-modified musculoskeletal questionnaire (Nordic Questionnaire). Chi-square tests and binary logistic regression were performed to analyze associations of MSD symptoms between study variables. 155 (58.9%) study participants reported MSD symptoms in past 12 months, in lower back (n = 126, 81.3%), leg (n = 122, 78.7%), neck (n = 106, 68.4%) and knee (n = 105, 67.7%); and 121 (46.1%) in last 7 days, in leg (n = 107, 88.4%), lower back (n = 102, 84.3%), neck (n = 89, 73.6%), upper back and knees (n = 87, 71.9%). Binary logistic regression showed relationship between working hours, seat and road condition, average load per ride, rickshaw ride duration, and standing driving with MSD symptoms among rickshaw drivers, especially in the lower back, leg/calf muscles, neck and knees. The results showed a high prevalence of MSD among all rickshaw drivers, with the neck, lower back, leg/calf muscles and knees being the most affected body parts. In order to avoid adverse effects on the occupational health of rickshaw drivers, ergonomic intervention training is necessary.