Background:  Hospital-acquired infections are the major concern of the health care system and agencies particularly in the isolation department. Nurses have the key and most important role in applying infection control measures in isolated departments. Aim of study: This study aimed to evaluate the effect of educational program on nurses' performance and compliance regarding infection control measures in isolated departments. Design: Quasi-experimental research design was utilized to conduct the aim of this study. Setting: The study was conducted in the isolation departments at Benha University Hospital. Sample: A convenient sample of all available nurses (60) from both sexes who are working at the previous mentioned setting. Tools: Three tools are used Tool I: A Self – administered questionnaire Tool II: Observational checklist Tool III: ـ The Isolation Precautions Compliance Scale. Results: 18 % of the studied nurses had satisfactory level of the total knowledge at preprogram implementation which improved to 85% immediately post and slightly decreased to  78.3% after 3 months  post program implementation. It showed statistically highly significant improvement between total practice and compliance immediately post and after 3 months of program implementation as compared with pre program p (0.001**). Conclusion: Educational program effectively improved the studied nurses' knowledge and practice regarding infection control measures with a highly statistical significance correlation between nurses performance and their compliance in the isolation department. Recommendation: Providing continuous educational or training program for nurses regarding occupational hazards and especially on protective measures.