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Purpose: The purpose of this study was to investigate occupational stress and fatigue level of neonatal nurses and to assess relationship between the two of them. Methods: Subjects were 109 neonatal nurses working at 2 university hospitals and 9 women's hospitals in D city, and the data were analyzed using descriptive statistics and binary logistic regression analysis by the SPSS/PC win 15.0. Results: The mean score of nurses` occupational stress level was 3.22, and the highest was 'night shift' among the 7 subcategories, and fatigue level was 62.0, and the highest was physical fatigue in configuration field. In logistic regression analyses, 'heavy workload’ and ‘inadequate physical environment’ led to most occupational stress. Conclusion: In addition to significantly higher levels of occupational stress of neonatal nurses, the cumulative fatigue also resulted in decreased quality of nursing, job satisfaction and nursing productivity; therefore, appropriate number of nurses and improvement of work environment is necessary. Furthermore, to reduce the occupational stress and fatigue, we should try to develop adequate clinical guidelines and intervention strategies and apply them in neonatal care unit.