〔Abstract〕 To explore the microbiological characteristics of Staphylococcus epidermidis with hemolytic phenotype (SEHP). Methods Hemolytic phenotype was detected using the three-point inoculation method, involving a total of 5 strains of SEHP and 5 strains of S.epidermidis with non-hemolytic phenotype (SENHP) . Bacterial species were identified using the Microflex LT MALDI-TOF mass spectrometer, and a phylogenetic tree was constructed through 16S rRNA sequence alignment. Antimicrobial susceptibility testing of SEHP and SENHP against commonly used antibiotics was performed using a VITEK 2 GP639 test kit. Antagonistic effects of SEHP and SENHP against Staphylococcus aureus and Corynebacterium striatum were evaluated by the Oxford cup inhibition assay. Red blood cell toxicity was detected using the microplate method. Biofilm formation ability was assessed by microplate crystal violet staining. Growth curves were monitored through the microcultivation assay. Results Compared with SENHP, SEHP had lower resistance rates to levofloxacin and moxifloxacin. Additionally, SEHP could antagonize S. aureus and C. striatum, exhibiting significant hemolytic toxicity, as well as a marked decrease in growth rate during the late logarithmic phase. Conclusion The microbiological characteristics of SEHP differ from those of SENHP in that SEHP demonstrates antagonistic effects against S. aureus and C. striatum, warranting further investigation.