〔Abstract〕 Objective To explore the microbiological characteristics ofStaphylococcus aureus(S.aureus) with novel incomplete hemolytic phenotype(SIHP). Methods Hemolytic phenotypes were detected and categorized by using the three-point inoculation method. A total of 11 novel SIHP and 33 randomly matchedS.aureuswith complete hemolytic phenotype(SCHP) were included. Antibiotic susceptibility test was performed using broth microdilution method. Coagulase test was performed with freeze-dried rabbit plasma. Catalase activity was detected by slide catalase test. Expression of hemolysin genes was detected by qRT-PCR. Toxicity to human red blood cells was assessed by microplate method. Microplate biofilm formation was measured using crystal violet staining method. Growth kinetic determination was performed through microcultivation assay. Results Compared with SCHP, the expression profiles of the four hemolysin genes(hla,hlb,hlc, andhld) in the new SIHP were different. The new SIHP had higher resistance rates to penicillin, oxacillin, gentamicin, quinolones, clindamycin, and trimethoprim-sulfamethoxazole. Furthermore, the new SIHP had stronger hemolytic toxicity, plasma coagulase activity, and biofilm formation ability. Additionally, the new SIHP grown faster in the logarithmic phase. Conclusion Taken together, the microbiological characteristics of the new SIHP are different from those of SCHP, including stronger antibiotic resistance and pathogenicity, which should be paid more attention by clinicians.