Purpose: The purpose of this study was to develop guidelines for cleaning of enteral nutrition bags by comparing the level of equipment contamination according to cleaning and disinfection methods. Method: This study was a true-experimental study, with 60 cases in total. Twenty cases each were randomly assigned to tepid water, detergent and brush, and disinfectant groups. The period of the experiment was March to April 2010, and enteral nutrition was given for 1 houre, 3 times a day at 7AM, noon, and 7PM for seven days. Enteral nutrition bags were cleaned after each feeding according to assigned cleaning and disinfection method followed by microbial cultures on 4th and 8th day before the 7AM feeding. Results: After 3 days of feeding and cleaning, the level of contamination of bags was not significantly different among the three groups. After seven days, the level of contamination was significantly lower when bags were cleaned with detergent and brush or with disinfectant compared to cleaning with tepid water. Conclusion: In cases where enteral nutrition bags are reused for example, in home care settings, studying findings indicate that cleaning bags using detergent and brush or disinfectant is an effective way to prevent contamination of bags.