Surgical site infections (SSIs) are one of the most important healthcare-associated infections. The SSIs play a major role in increasing morbidity and mortality rates, prolonging hospitalization, and increasing hospital readmissions. The aim of the study was to assess nurses' knowledge and practice regarding wound infection in surgery unit at Assuit University Children s' Hospital. A descriptive research design was used in this study. Subjects and Methods: the study was carried in surgery unit of Assuit University Children Hospital. A convenience sample of 30 nurses working in surgery unit of Assuit University Children Hospital. Data collection tools were a structured questionnaire interview sheet to assess nurses' knowledge and an observation checklist to evaluate nurses' practice regarding wound infection. Results were shown satisfactory level of knowledge and unsatisfactory level of practice regarding wound infection in surgery unit at Assuit University Children Hospital. The study concluded that nurses had satisfactory level of knowledge and deficiency in practice. Statistically significant relations were shown between knowledge and practices among studied nurses. The study recommended Provision of continuing education programs associated with clinical training related to infection control