Objective: To determine the nurses’ knowledge toward prevention of sepsis in neonatal care unit in Mosul city. Methodology: This descriptive hospital-based study included nurses from pediatric teaching hospitals in Mosul city from 10th March 2022 to the 15th May 2022. The sample size consisted of all available (60) nurses, who worked in the hospital during the study period. Data were collected using a questionnaire designed by the researchers. Data was analyzed by SPSS version 25. Results: The majority of the study sample were females (68.3%) and 31.7% were male. According to age, 40% of the sample was aged between 20-24 years, while 31.7% age ranged between 25-29 years. Majority of the sample’s level of education was diploma (36.7%), while 20% were bachelor. A third of the study sample (31.7%) had no previous training related to sepsis. Conclusion: In a third of the study population, no prior training in sepsis and its prophylaxis was available. Nurses at the neonatal care unit in Mosul City had good knowledge about sepsis prevention, and their educational background, length of hospital employment, years of experience, and training program all had an impact. There are no statistically significant differences in the expertise of nurses based on their gender or age.