Purpose: The purpose of this study was to examine the influence of a handrail (presence and position) on treadmill gait and balance in stroke patients during gait training. Methods: 39 patients with stroke (male 31, female 8) participated in this study. The training groups were classified into a no-handrail group (NHG), front handrail group (FHG), and bilateral handrail group (BHG). Each group comprised 13 subjects. The subjects were trained to walk in a straight path 30 minutes per day for 8 weeks. The Good Balance System was used to measure static balance and dynamic balance. To measure walking ability, timed up and go (TUG) was also assessed. Results: The NHG showed no significant differences in static balance, dynamic balance, and TUG. The FHG was significantly different in their medial-lateral speed of static balance, dynamic balance, and TUG. The BHG was significantly different in their static balance, dynamic balance, and TUG. Conclusion: These findings consider the effects of holding handrails concomitantly with changes in postural stability. We conclude that for training stroke patients, treadmill walking while holding handrails improves balance and gait more than treadmill walking without holding handrails. The resulting changes in muscle activity patterns may facilitate the transfer to a gait pattern. The results of this study suggest methods for training treadmill walking in stroke patients.