Background : The retained surgical item is a critical double burden to the patients and their families. One way of minimizing the risks of this critical burden is the surgical counting process which is costless, effective and preventive method. The present study aimed to determine the knowledge and practice towards surgical counting among operating room nurses and midwives at referral teaching hospitals in Rwanda. Methods: A descriptive cross-sectional study was conducted at four referral teaching hospitals on 160 randomly selected nurses and midwives working in the operating rooms. Nurses and midwives working in OR reported their knowledge and practices in surgical counting exercise using a self-administered questionnaire. A checklist was used by the researchers to observe the compliance with surgical count policy in the OR. Data were entered in Statistical Package for Social Sciences (SPSS version 21.0) for analysis. A correlation between knowledge and practices and association between demographic characteristics with knowledge and practices were tested in this study. The ethical principles were valued. Results: Out of 156 participants who responded to this study, the majority were females (62.8%), Married (70.5%), 30-39years old (53.8%), advanced diploma (82.1%) and with clinical experience below 6 years (52.5%). The majority (78.2%) had good knowledge of surgical count but the majority reported the poor practices (57.8%, n=89). Compared to self reported practices, the observation reports of 35 cases revealed very poor performance. Being female and 1-3 years experience in OR are factors for performing surgical counting practices (OR:3.030562;p≤0.027); and (OR; 9.215279;p ≤0.002) respectively). Conclusion : The surgical count practices were self-reported and observed at low level. The experience and female gender was associated with the practices towards surgical count safety. Others barriers that may hinder the practices were not explored in this study and recommended for further research. Increasing a number of specialized perioperative nurses in OR and in service training and policy implementation follow up may improve Surgical count safety practices. Key words: Surgical count, Operating room, Practice and Referral hospital