Objective To explore the application of the Handbook for the Training Nurses in the standardized training for nurses in pediatric surgery. Methods Twenty-four training nurses trained under the instruction of the Handbook for the Training Nurses in Department of Pediatric Surgery from July 2014 to December 2015 were included as the observation group. Their training effects were compared historically with those of twenty-four training nurses who were trained without the help of the handbook from January 2013 to June 2014 (the control group). Results After the completion of the half-year training, theoretical score of training nurses in the observation group was better than that in the control group, and the difference was significant (87.16±4.18 vs. 83.71±5.46; t=2.492, P=0.016). Meanwhile, the specialist practical skill examination of training nurses in the observation group, such as femoral vein blood collection, replacement of drainage bag and indwelling needle infusion in children, and emergency practical skill examination such as cardiopulmonary resuscitation and the use of simple respirator were higher than those in the control group; and the working performance scores of training nurses in the observation group, such as the ability to adapt to the environment, patient satisfaction, nursing document writing, work efficiency and morning questions were better than those in the control group; the differences were significant (P< 0.05). Conclusion The Handbook for the Training Nurses can help the training nurses to adjust their working environment, master the practical skills and nursing knowledge in pediatric surgery.