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Background & Aims: Failure in proper endotracheal suctioning may lead to hemorrhage, lesions in the tracheal mucosa, infections, atelectasis, and hypoxia, which are associated with increased mortality, length of hospital stay, and treatment costs in hospitalized neonates. Considering that nurses play a key role in the prevention of these complications, the present study aimed to assess the effect of training on the performance of nurses in the endotracheal suctioning of neonates. Materials & Methods: This quasi-experimental was conducted with control groups on 80 nurses, who were selected via continuous sampling, at selected hospitals affiliated to Tehran University of Medical Sciences and Iran University of Medical Sciences (two control hospitals and two intervention hospitals) in 2016. Before the training intervention, the performance of nurses was evaluated using a researcher-made checklist, which was developed with a demographic section and nursing performance section with 30 items based on guidelines and expert opinions. The contents of the checklist and educational contents were assessed and approved by a panel of experts. The performance of nurses was observed by the researcher in three stages. One month after the intervention, the performance of the same nurses was evaluated again by the same researcher in three stages. Results: The score of nursing performance before and after the intervention was 51.38±8.70 and 72.19±4.16, respectively, which indicated a statistically significant difference (P |