Purpose: To develop a web-based program on blood-borne infection control and to examine the effect of the newly developed program on perceived threat of diseases, knowledge, preventive health behaviors for blood-borne infections, and incidence rates of accidental needle sticks and other sharp object injuries in nurses. Methods: The program was developed through the processes of analysis, design, development, implementation, and evaluation. The research design involved a nonequivalent control group for pretest and posttest experiments. The setting was a 745-bed general hospital located in Korea. Results: The program was designed and developed after consulting previous studies. After development of the program was completed, it was evaluated and revised by a panel of experts. The total score for perceived threat of diseases, knowledge, preventive health behaviors in the experimental group was significantly higher compared to the control group (p<.05). The incidence rates for needle sticks and other sharp object injuries in the experimental group were significantly lower compared to the control group (p<.05). Conclusion: Application of a Web-based, blood-borne infection control program is effective, and can be expanded to other healthcare workers who also have a high risk of blood-borne infections.