耐万古霉素屎肠球菌基因检测与多位点序列分析分型研究  

Drug resistance genes in vancomycin-resistant Enterococcus faecium and MLST genotyping

作　　者：燕吉广[1] 董春忠[1] 赵广敏[1] 周卫岭[1] 孙霞 王月玲[3] 

机构地区：[1]滨州市人民医院,山东滨州256610 [2]滨州市妇幼保健院,山东滨州256610 [3]山东大学附属省立医院检验科,山东济南250021

出　　处：《中华医院感染学杂志》2016年第18期4092-4094,4097,共4页Chinese Journal of Nosocomiology

基　　金：山东省医药卫生科技发展计划基金资助项目(2014WS0081)

摘　　要：目的了解医院分离的屎肠球菌耐药基因分布及MLST分型特点,为医院合理应用抗菌药物和医院感染控制提供理论依据。方法收集医院2013年1月-2014年12月分离的肠球菌112株,采用Phoenix微生物全自动鉴定系统进行菌株鉴定;采用PCR方法检测Van A、Van B、Van C1、Van C2/3、Van D、Van E、Van G、VanM、TetM、ermB、TEM、aac(6′)-aph(2")、aph(3′)-Ⅲ、ant(6′)-Ⅰ等耐药基因;扩增7个管家基因adk、atpA、ddl、gdh、purK、gyd、pstS确定屎肠球菌的序列型。结果 3株耐万古霉素屎肠球菌2株Van A、1株Van B、3株ermB、3株aac(6′)-aph(2")、3株aph(3′)-Ⅲ基因阳性;MLST分型为3个ST型,1号菌株为ST78型,2号菌株、3号菌株为新的ST型分别是ST1014、ST1015;药敏试验显示,万古霉素MIC≥256μg/ml、替考拉宁的MIC≥256μg/ml或MIC 16μg/ml、对利奈唑胺MIC 2μg/ml。结论耐万古霉素屎肠球菌不断增多,医护人员应加强手卫生、接触隔离、环境消毒、合理使用抗菌药物等措施,以控制VRE的感染发生。OBJECTIVE To understand the drug resistance genes in clinical Enterococcus faeciumisolates and observe the characteristics of MLST genotyping so as to provide theoretical basis for reasonable use of antibiotics and control of nosocomial infections.METHODS A total of 112 strains of Enterococcus were isolated from Jan 2013 to Dec 2014,the strains were identified by using Phoenix microorganism automatic identification system,the drug resistance genes,including Van A,Van B,Van C1,Van C2/3,Van D,Van E,Van G,VanM,TetM,ermB,TEM、aac（6′）-aph（2 ＂）,aph（3′）-Ⅲ,and ant（6′）-Ⅰ,were detected by means of PCR;7housekeeping genes,namely adk,atpA,ddl,gdh,purK,gyd,and pstS,were amplified so as to determine the sequences of the E.faecium.RESULTS Of 3strains of vancomycin-resistant E.faecium,2were detected positive for Van A,1strain positive for Van B,3strains positive for ermB,3positive for aac（6′）-aph（2＂）,3positive for aph（3′）-Ⅲ.Totally 3types of ST were identified through the MLST genotyping,the NO.1strain was ST78,NO.2strain and NO.3strain were the novel ST types,which were ST1014 and ST1015,respectively.The drug susceptibility testing showed that the MIC of vancomycin was no less than 256μg/ml,the MIC of teicoplanin no less than 256μg/ml or 16μg/ml,the MIC of linezolid 2μg/ml.CONCLUSIONThe vancomycin-resistant E.faeciumstrains are increased;it is necessary for health care workers to take measures such as hand hygine,contact isolation,environmental disinfection,and reasonable use of antibiotics to control of VRE infection.

关 键 词：屎肠球菌 万古霉素 耐药 VAN A VAN B MLST分型 

分 类 号：R378[医药卫生—病原生物学]