Staphylococcus aureus (S. aureus) is one of the common pathogens causing healthcare-associated infection (HAI) and community-associated infection (CAI). In recent years, there are more and more failure cases in the anti-infection treatment of S. aureus in clinical practice, and the formation of biofilm is considered the main cause for the failure of antimicrobial treatment. However, drug resistance mechanism of S. aureus biofilm has not been fully elucidated. Evidence shows that S. aureus biofilm infection is difficult to cure and easy to reoccur, and repea-ted treatment after infection greatly increases the pain and economic burden of patients. In this paper, research progress of drug resistance mechanism of S. aureus biofilm is reviewed, so as to provide reference for the development of new antimicrobial agents.