Objective To analyse the resistance and horizontal gene transfer of high level erythromycin resistant Enterococcus faecium.Method PCR technique was applied to identify genes including ermB,mef,and tnpA.Plasmid DNA was treated with restriction enzymes and subjected to agarose gel electrophoresis for the analysis of DNA fragments.The transferability of the erythromycin resistance was examined by solid mating and broth mating.Result Of the 90 HLER E.faecium,85.6%(77/90) were positive for ermB gene,and 15.6%(14/90) positive for both tnpA and ermB gene. All isolates were negative for mef gene. Majority of the plasmid patterns were identified in single isolates. 14. 3% (11 /77) isolates with ermB were horizontal transferred between E.faecium strains in solid mating. 2.6% (2/77) isolateswere horizontal transferred between E.faecium in broth mating. The 13 conjugations were positive for ermB gene, and negative for tnpA gene. Conclusion This study indicated that the positive rate of ermB gene were higher among HLER E.faecium isolates and this gene can horizontal transfer between E.faecium. The ermB gene which can horizontal transfermaybe on other transposon Tn917 and /or conjugative p lasmids. The ermB gene of the 14 HLER E.faecium maybe on Tn917. There were no obvious homology in the HLER E.faecium isolates.