OBJECTIVE: To investigate the knowledge of rational use of antibiotics among pharmacists in medical institutions in China’s Shanxi province, so as to find out the problems, then to provide support for the management of antibiotics. METHODS: A questionnaire survey was conducted, which included the basic information of the respondents, the basic knowledge of antimicrobial management and the related knowledge of antimicrobial. SPSS 25.0 was used for statistical analysis. RESULTS: 462 pharmacists were investigated. The average score of the knowledge related to rational use of antibiotics was (10.49±4.05). It showed that the hospital type, grade, pharmacist’s education, professional title and working years all had effect on the pharmacists’ mastery of antimicrobial related knowledge (P<0.05). Multivariate logistic regression analysis showed that hospital grade and pharmacist’s education were the main influencing factors (P<0.05). CONCLUSION: Pharmacists have insufficient knowledge about the rational application of antibacterial drugs. It is essential to strengthen the training of management regulations and application of antibacterial drugs.