Abstract: OBJECTIVE To explore the intervention effect of linezolid combined with vancomycin on biofilms of clinical methicillin-resistant Staphylococcus aureus（MRSA） isolates. METHODS Totally 120 strains of MRSA were isolated from Wuhan No.1 Hospital between Jan 2020 and Jan 2021. The minimum inhibitory concentrations（MICs） and fractional inhibitory concentrations（FICs） of vancomycin and linezolid against MRSA were determined, the time-killing curves were drawn, and the effects of linezolid and vancomycin alone and in combination on the biofilm of MRSA were observed by crystal violet staining method and electron microscope scanning. The relative expression levels of biofilm-related genes（icaA, icaD, cidA, agrA and sarA）mRNA in MRSA were detected by real-time fluorescent quantitative PCR（RT-qPCR）. RESULTS The MICs of both linezolid and vancomycin ranged between 0.5 and 2 μg/ml, the MIC of linezolid combined with vancomycin ranged between 0.125 and 1 μg/ml, and the MIC, MIC50 and MIC90 of the combination of the two drugs were less than those of the single use（P＜0.05）. The synergistic effect（73.33%） and additive effect（22.5%） were dominant among the combination of the two drugs, few showed irrelevant effect（4.17%）, and no antagonistic effect was observed. The combined and single use of linezolid and vancomycin showed remarkable antibacterial effect on MRSA after intervention for 24 hours; the OD₅₅₀ of the combined use was lower than that of the single use（P＜0.05）. The formation of MRSA biofilms was inhibited, which could be seen under scanning electron microscopy. The relative expression levels of MRSA biofilm formation-related genes icaA, cidA, agrA and sarA mRNA were downregulated when the two drugs were used in combination or alone（P＜0.05）, and the effect of the combined use was greater than that of the single use（P＜0.05）. CONCLUSION Linezolid and vancomycin show synergistic or additive effect on inhibition of MRSA, the combination of the two drugs shows more remarkable effect, and the mechanisms may be associated with the expression of mRNA of genes that are related to inhibition of MRSA biofilms.