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Purpose: The purpose of this study was to confirm the relationship between aseptic technique performance in clinical nursing skills and nurse's role overload and to provide the basic data to decrease the incidence rate of nosocomial infection. Method: Participants(N=283) were recruited in B city from April 2007 to June 2007. The data were collected by a structured questionnaire and analyzed with descriptive statistics, t-test, ANOVA and Pearson's correlation coefficient. Results: The level of aseptic technique performance as basis for insertion of foley catheterization with the participants was 42.72 out of the total score, 50; that of as basis for insertion of intravenous catheter for fluid therapy was 40.11 out of the total score, 55. The level of the nurse's role overload was 9.85 out of the total score, 15. There was a significantly positive relationship between the aseptic technique performance in insertion of foley catheter and that of intravenous catheter for fluid therapy(r= .279, p<.001). There was not a significant relationship between aseptic technique performance and nurse's role overload. Conclusion: Based on the finding, we could conclude that nurse's duty of attention for asepsis is strictly needed in performing nosocomial infection-related clinical nursing skills.