Objective: To assess the knowledge, attitudes and practices of providers in blood transfusion, HIV infections and hepatitis B and C in the province of South Kivu (eastern DRC). Methods: This study is comparative cross-sectional. It covered all the staff care. Our sample was composed by doctors, nurses, midwives, and agents’ rapid training in activity in health sector in South Kivu between 1 December 2011 and 30 December 2011. Those medical and paramedical staffs came from 12 general reference hospitals, 11 hospitals and 65 health centers in the province of South Kivu. Three hundred and ninety three people were interviewed including 6.5% of physicians, 87.4% of nurses and 5.1% of midwives. Among them, 25.5% had received training in blood transfusion. Results: Overall, 11.7% of providers responded correctly to questions on knowledge and attitude on blood safety. The proportion of correct answers on the knowledge and attitude on exposure accidents was 53.8%. The frequency of caregivers who responded well to the basics of HIV and hepatitis B and C was 15.5%. Notions about the type of donor blood and the risk of blood transfusion received an overall rating of 17.8% and the proportion of correct answers on the knowledge and practice base of blood transfusion was 16.7%. During the six months following the survey, 54.4% of providers have experienced accidental exposure to blood. Conclusion: The lack of knowledge of the medical staff in transfusion is obvious. We must therefore have the possibility of organizing regular training and follow-up activities.