RFID technology, an automatic identification and data capture technology to provide identification, tracing, security and so on, was widely applied to healthcare industry in these years. Employing HEPA ventilation system in hospital is a way to ensure healthful indoor air quality to protect patients and healthcare workers against hospital-acquired infections. However, the system consumes lots of electricity which cost a lot. This study aims to apply the RFID technology to offer a unique medical staff and patient identification, and reacting HEPA air ventilation system in order to reduce the cost, save energy and prevent the prevalence of hospital-acquired infection. The system, reacting HEPA air ventilation system, contains RFID tags (for medical staffs and patients), sensor, and reacting system which receives the information regarding the number of medical staff and the status of the surgery, and controls the air volume of the HEPA air ventilation system accordingly. A pilot program was carried out in a unit of operation rooms of a medical center with 1,500 beds located in central Taiwan from Jan to Aug 2010. The results found the air ventilation system was able to function much more efficiently with less energy consumed. Furthermore, the indoor air quality could still keep qualified and hospital-acquired infection or other occupational diseases could be prevented.