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Background: Prescribers must possess extensive knowledge and maintain a positive attitude towards the rational use of medicines to achieve desirable treatment outcomes and effectively prevent treatment failures, increased costs, drug toxicities, and interactions. The objective of this study was to evaluate prescribers' understanding and perception concerning the rational use of medicines in public hospitals. Additionally, the study aimed to identify the factors that influence rational prescribing practices., Methods: A structured data instrument was developed to collect demographic data and evaluate participants' knowledge and perception of rational medicine use, in line with the study objectives. Chi-squared statistics and Fisher's exact test were utilized to identify factors associated with good knowledge and perception among participants. Logistic regression was then employed to assess the strength of the associations, with odd ratios reported at a significant level of 0.05., Results: Out of 192 participants, 85.4% held a positive view of rational medicine use, stressing patient safety and recognizing risks like antimicrobial resistance and polypharmacy. Perception was influenced by factors such as prescriber profession, access to references, and drug bulletin updates. Additionally, 65.6% demonstrated good knowledge of rational medicine use, which was notably influenced by factors like using standard prescribing guidelines, having a functional Drug and Therapeutics Committee, prescriber profession, and the frequency of drug bulletin updates., Conclusion: The study emphasizes the critical need to address knowledge gaps among healthcare professionals, especially nurses and other prescribers, to ensure the safe and effective use of medications. It highlights the positive influence of utilizing preferred prescribing references and the existence of functional Drug and Therapeutics Committees in hospitals on knowledge levels. However, the unexpected findings regarding the limited impact of frequent updates of drug bulletins require further investigation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Djochie et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)