Objectives This study aim was to evaluate the effectiveness of nurses training program on knowledge, and practice of nosocomial infection control at Cho Ray hospital in Vietnam. Method Pretest and posttest study design was used. A total of 108 nurses derived from 6 departments at the HepatoBiliary- Pancreatic, Orthopaedics, Digestive, Burns, Neurology A and Neurology B of Cho Ray Hospital participating in this research since 15/10/2014 to 15/02/2015. A survey on nurses’ knowledge of nosocomial infection control as a baseline measurement, followed by 6 week training program as intervention strategy, the statistical differences were used to analyze the effectiveness of the training program derived from the baseline measurement and post-test. Finally, a checklist was used as a to observation tool to evaluate nurses’ hand washing, and intravenous injection skills. Results There was a significant improvement in knowledge of hospital infection control among 108 nurses participating in this research. Before the training program, only (2.8%) 3/108 of the nurses could answer correctly all of 26 questions about general knowledge of infection control. However, 58.1% (63/108) of nurses answered all the questions correctly after six weeks training program. A paired t-test was used which indicated that a statistically significant difference between baseline and post-test after training with p <0.01. Training programs enhance nurses’ knowledge of infection control demonstrated training effect. The nurses also improved practice on hand hygiene which according to guidelines recommended by WHO and Ministry of Health of Vietnam, and intravenous injection skills significantly. Conclusion From results of this study implied that it was necessary to organize a standard training program of nosocomial infection control periodically for nurses to update their knowledge and practice. Even though they were well trained before working for the hospital, without regularly reminded about the importance of preventing nosocomial infection, their knowledge and quality of practice on nosocomial infection might be deteriorated over time.