Introduction: Premature infant had a greater chance of complications which lead to death. The world health organization (WHO) reported that out of 130 million live births every year, 4 million die within the first four weeks of life. Of these deaths 99% occur in developing countries against 1% in developed countries. Objective: This study aimed at assessing the effect of training program on nurses’ practice regarding premature nursing care in Pediatric Teaching Hospital. Methodology: This is an interventional hospital based study including all available nurses (101) during the period of the study from (February to April 2017). Results: The study results evident that Among of 101 of nurses participate in the present study was show: that: that: 40.6% of the study sample at the level of bachelor’s degree, 19.8% of them their years of experience ranged from 6 – 10 years & 14.9% of the study sample attended training program before(Figure1,2&3). The mean of the total percentages of nurses’ performance regarding maintenance of temperature of premature infant was 39.6% before the training program and it was improved to 84.1% after the program (4.10). The mean of the total percentages of nurses’ performance regarding maintenance of fluid of premature infant was 33.4% before the training program raised to91.4 % after (4.11). The mean of the total percentages of nurses’ performance regarding skin care of premature infant was 25.8% before the training program improved to 85.1% after (4.12). The mean of the total percentages of nurses’ performance regarding maintenance of nutrition of premature infant was 21% before the training program and it was improved to 79.4 % after the program (4.13). The mean of the total percentages of nurses’ performance regarding prevention of neonatal infection was 29.5% before the training program and it was improved to 80.3 % after the program (4.14).