Background: Hand hygiene is one of the effective measures for infection prevention and rncontrol. HCAIs are the primary causes of illness and death in NICUs. It has been estimated thatrn30-40% of HCAIs are related to contamination of HCW hands especially nurses have a contactrnfor a long time with neonates. Thus, this study is intended to provide information on the level ofrnknowledge, attitude, and practice of hand hygiene among NICU nurses. rnObjective: To assess knowledge, attitude, and practice of hand hygiene among NICU nursesrnworking in public hospitals, Addis Ababa, Ethiopia, 2020. rnMaterials and methods: Institutional based cross-sectional study design was conducted andrnpurposive sampling was used for selecting hospitals. All 120 NICU nurses working in five public rnhospitals under AAHB from February 18 to March 25, 2020, were included in this study. Datarnwere collected through a self-administered questionnaire and observational checklist whichrnconsists of four sections. A pretest was carried out in TASH and data was collected by trainedrnhealth professionalsâ€™ under the supervision of a trained supervisor. The collected data werernentered using Epi Data version 4.6.0.2 and analyzed using SPSS version 25 software. Descriptivernstatistics (frequency, percentage, and mean) and bivariate logistic regression analysis were used.rnFinal results were presented using texts, pie-charts, tables, and graphs. rnResults: A total of 114 NICU nurses with a response rate of 95% have participated. This studyrnshowed that the majority of the NICU nurses 96(84.2%) had moderate, 15 (13.2%) had good and rn3(2.6%) had poor knowledge. Level of attitude majority of nurses 88(77.2%) had moderate,rn23(20.2%) had good, and 3(2.6%) had poor. Most of them 93(81.6%) had poor, 21(18.4%) hadrnmoderate and 00(0.0%) had a good practice on hand hygiene. There was no significantrnassociation between dependent variables (KAP) and socio-demographic variables. rnConclusions and recommendations: Based on the study findings it can be concluded thatrnmajority of them had moderate knowledge and attitude but they had poor practice regarding hand rnhygiene. Therefore, it is recommended to give training on hand hygiene guidelines includingrnmonitoring the practice regularly.