ObjectiveTo investigate the awareness of knowledge, attitude and behavior about disinfection and sterilization among clinical health care workers (HCWs). Methods200 clinical HCWs were investigated randomly by using questionnaires.ResultsThe average awareness rates of knowledge, attitude and behavior about disinfection and sterilization among HCWs were 71.49%, 63.79%, and 86.45% respectively.  The awareness rates of knowledge about the expiration dates of sterilized articles, classification of infectious diseases, and biological monitoring were 49.43%，42.53%，  and  23.56%, respectively； HCWs’ attitude towards disinfection and sterilization：awareness rate of whether they paid attention to the new technology in specialty of central sterile supply was the lowest (26.44%) ; 86.45% of HCWs thought they could master the aseptic operation technique, correct application method for sterile apparatus and related items，but easily ignored to check the integrity of aseptic packages and indicators before operation (67.24%).ConclusionThe knowledge, attitude and behavior about disinfection and sterilization among HCWs still need to be improved，training on knowledge about disinfection and sterilization among HCWs should be strengthened.