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The purpose of this study is to develop nursing standard guidelines for nurses in a neonatal intensive care unit. The Delphi method was used in this study to elicit expert consensus. Thirteen experts who were nurses and pediatric adolescent specialists working in the neonatal intensive care unit participated in the study. In this study, 178 items were developed based on 5 nursing practice standards and 7 standards of professional practice. An additional 10 items were included based on observation in the neonatal intensive care unit. After expert validation, a final total of 184 items was developed. The standard guidelines for high-risk neonatal care developed in this study for practical clinical education in nursing are significant because they reflect the nursing practice standards in Korea and characteristics of nursing practice in the neonatal intensive unit.