Objective To investigate the drug resistance phenotype, drug resistance genes and molecular characteristics of clinical isolates of Staphylococcus aureus in Yangzhou from 2016 to 2020. Methods A total of 71 strains of clindamycin resistant Staphylococcus aureus were collected. Molecular typing of the strains was performed using multilocus sequence typing (MLST) and SNP. Antimicrobial minimal inhibitory concentrations were detected by the method of microdilution broth. Different resistance genes were detected. Results A total of 71 strains of clindamycin resistant Staphylococcus aureus were collected in this study, of which 42 strains (59.15%) were inherently resistant and 29 strains (40.85%) were induced resistant ; 44 strains (61.97%) were found to be methicillin resistant Staphylococcus aureus (MRSA). Among the 71 strains , 18 ST types were found , and the detection rates of mecA and ermB in ST59 strain were higher than those in other ST types. Among the clustered strains, CC59 was inherently resistant. Conclusion The clinically isolated Staphylococcus aureus in Yangzhou has severe drug resistance, especially the resistance to clindamycin. CC59 clone group carries many drug resistance genes, and all of them are MRSA . This study provides reliable data for clinical selection of appropriate drugs and further investigation of the prevalence of clindamycin-resistant strains.