Purpose: The purpose of this study was to develop a web-based education program on control vancomycin-resistant enterococci (VRE) infections and to identify the effects of the program on knowledge and performance of nursing students. Methods: The web-based VRE infection control education program was developed by using the network-based instructional systems design model. The nursing students in the experimental group could access this web-based education program at any time, and as many times as they wanted, during the clinical training period. Effects were evaluated by assessing knowledge and performance of VRE infection control measures during the clinical training period. Results: The contents of the education program included diagnosis, transmission, and treatment of VRE, contact precautions, hand washing, personal protective equipment, environment management, and quizzes. The lecture portion was filmed in a virtual screen studio using flash animation, video, and sound effects, and it was uploaded on an internet site. The knowledge and performance scores of the experimental group after using the education program were significantly higher than those of the control group. Conclusion: The results suggest that the web-based VRE infection control education program is an effective educational method to enhance knowledge and performance of VRE infection control measures.