Background: Glucose -6- phosphate dehydrogenase (G6PD) deficiency is one of the most common genetic enzymes abnormalities leading to hemolytic anemia affecting more than 400 million children worldwide. Aim: this study aimed to assess nurses' knowledge and performance related to care of children with glucose-6- phosphate dyhydrogenase deficiency at Assiut Children University Hospital. Design: Descriptive research design was used in this study. Subjects and Method: The subjects who participated in this study included all nurses (45) working in the General Hematology unit (18),Private Hematology unit (8) and Emergency unit (19) at Assiut University Children's Hospital.Tools: a structured interview sheet and observational checklist were used to assess nurses' knowledge and performance. Results: It was found that more than half of nurses had unsatisfactory total score of knowledge and performance related to care of the children with G6PD deficiency. There were statistical significant relation between total score of nurses' knowledge, performance and their personal characteristics except the residence and place of work. Conclusion: Nurses knowledge regarding G6PD deficiency were not enough with some unsafe practices. Recommendation: Continuing nursing education and in-services training programs in general, special hematology and emergency units concerning glucose-6- phosphate deficiency have to be emphasized periodically