Background: Assessment and control of nosocomial infections is a global priority which decreases length of hospitalization and also significantly reduces the health care costs by minimizing infections. So the present study evaluated the knowledge, self-efficacy and practice of nursing staff in Disciplines Standard Precautions to prevent nosocomial infection.Materials and Methods: This Cross-sectional and analytical study carried out on 130 nursing staff of hospitals of Arak university of medical sciences in 2013. Data collected with a valid and reliable questionnaire including of knowledge, self-efficacy and practice of nursing staff in Disciplines Standard Precautions. Data was analyzed by SPSS software using descriptive and analytical statistical methods and pearson correlation.Results: The mean score of knowledge was 46.92±14.66, practice 49.46±6.96 and average of self-efficacy was 52.2±10.81. There was not significant relationship between knowledge and Practice, but there was a significant relationship between self-efficacy, knowledge, practice and marriage (p<0.05). Moreover there was a significant relationship between knowledge and gender. (p<0.05).Conclusion: Despite moderate self-efficacy, majority of nurses did not have appropriate knowledge about prevention of nosocomial infections based on the findings of this study. Considering the important role of nurses in prevention of nosocomial infections, training is necessary to increase the prevention behaviors in nurses. Therefore, paying more attention to apply actions for training of nurses is necessary.