OBJECTIVE The in vitro activity of home-made rifamycin was investigated.METHODS Minimal inhibitory concentrations(MICs) were determined by agar dilution method using multipoint inoculator.RESULTS The results indicated that MIC50 and MIC90 of rifamycin against MRSA and MSSA were similar to vancomycin. The values were 1 and 2 mg／L, respectively. Rifamycin against MRSE was 4 times stronger than vancomycin.The value of MIC50 was 0.5 mg／L.The MIC50 of rifamycin against S.pneumoniae,Branhamella catarrhalis,and Listeria monocytogenes were 8,128 and 4 times lower than vancomycin, respectively.CONCLUSIONS There are good antibacterial activity of home-made rifamycin against meticillin resistant Staphyloccocus and the most species of clinical isolates,so can be used the infective diseases by MRSA and MRSE.