Introduction: The aim of this study was to determine mothers’ level of knowledge about dirty hand diseases and the practice of hand hygiene in the Tshopo health zone. Methods: A descriptive cross-sectional study was carried out among 200 mothers/caregivers selected by a three-stage cluster sampling technique, in the Tshopo health zone, using a structured questionnaire. Descriptive statistics were based on proportions, mean and confidence intervals. Results: The most frequently cited dirty-hand diseases were diarrhea (55%), typhoid fever (48%) and amoebiasis (41%). Inadequate hand hygiene (71.5%), soiled food (34%) and unsanitary water (21%) were the most frequently cited contamination routes. Hand hygiene was the best-known measure for preventing dirty hands (71%), and the most frequently cited times for practicing hand hygiene were before eating (77.5%) and after using the toilet (71%). Hand hygiene was most frequently observed before eating (66.5%) and after using the toilet (49%). Hand-washing facilities were close to toilets in 37% of cases. Conclusion: The knowledge of diseases of dirty hands and means of prevention is incomplete but that of hand hygiene as a means of prevention of these diseases is satisfactory. Raising awareness, the availability of washbasins, water and soap near the toilets are actions to be reinforced.