携带黏附素、杀白细胞素、α溶血素基因的MRSA耐药基因检测与样本聚类分析  

Detection of resistance genes and sample cluster analysis of methicillin-resistant Staphylococcus aureus carrying adhesion,leucocidin,andα-hemolysin

作　　者：程运涛 常青 吴为民 翁幸鐾[3] 陈国千[4] 王春新[4] 欧阳冰 

机构地区：[1]无锡市第八人民医院检验科,江苏无锡214000 [2]无锡市第八人民医院儿内科,江苏无锡214000 [3]宁波市第一医院检验科,浙江宁波315010 [4]无锡市人民医院检验科,江苏无锡214000

出　　处：《中华医院感染学杂志》2015年第19期4329-4331,4335,共4页Chinese Journal of Nosocomiology

基　　金：浙江省中医药基金资助项目(2011ZB126)

摘　　要：目的：了解携带黏附素、杀白细胞素、α溶血素基因的耐甲氧西林金黄色葡萄球菌（MRSA），对抗菌药物和消毒制剂耐药基因的存在及该组菌株的亲缘性。方法收集2012年1月－2013年7月儿童伤口标本中分离到的16株MRSA ，采用金黄色葡萄球菌蛋白A基因（spa）确定菌种，以苯唑西林耐药判定为MRSA ；采用聚合酶链反应（PCR）方法确定黏附素基因（sasX）、杀白细胞素基因（pvl）、α溶血素基因（hla）均为阳性，并采用PCR方法分析β‐内酰胺类、氨基糖苷类、大环内酯类、四环素类、季胺类消毒剂以及莫匹罗星等12种耐药基因，最后作基因检测结果的样本聚类分析。结果16株M RS A均携带黏附素、杀白细胞素、α溶血素基因，β‐内酰胺类、氨基糖苷类、大环内酯类、四环素类、季胺类消毒剂每一类耐药基因均有检出，且β‐内酰胺类耐药基因 mecA、氨基糖苷类耐药基因 aac（6′）／aph（2″）与 aph（3′）‐Ⅲ、大环内酯类耐药基因中 ermB、四环素类耐药基因 tetM检出率均为100．0％，未检出莫匹罗星耐药基因；16株MRSA耐药基因检测结果中，mecA＋ aac（6′）／aph（2″）＋ aph（3′）‐Ⅲ＋ ermB＋ tetM＋ qacA阳性模式共检出13株占81．3％；样本聚类分析提示，1～4，6、7、10～16号等13株为同一克隆，8号与9号亦为同一克隆；16株MRSA与MRSA‐ref亲缘关系较近；与MSSA‐ref亲缘关系较远。结论该组M RS A均携带黏附素、杀白细胞素、α溶血素基因，致病力很强，其对10种抗菌药物完全耐药，与耐药基因的高检出率一致，但莫匹罗星对该组菌株仍有抑制作用。OBJECTIVE To investigate distribution of resistance genes to antimicrobials and disinfections in a group of methicillin‐resistant Staphylococcus aureus （MRSA） carrying adhesion ,leucocidin ,α‐hemolysin ,and their rela‐tionships .METHODS Totally 16 strains of MRSA from wounds of children in the hospital from Jan .2012 to Jul . 2013 were collected ,and the S .aureus protein A ,spa ,was used to identify S .aureus .Oxacillin was used to iden‐tify MRSA ,adhesion gene （sasX） ,leucocidin （pvl） ,α‐hemolysin （hla） were all confirmed with PCR .Then ,12 kinds of resistance genes to beta‐lactams ,aminoglycosides ,macrolides ,tetracyclines ,quaternary ammonium disin‐fectants and mupirocin were used to perform the sample cluster analysis of the detection results of resistance genes .RESULTS Adhesion ,leucocidin ,α‐hemolysin as well as each class of resistance genes to beta‐lactams ,ami‐noglycosides ,macrolides ,tetracyclines ,and quaternary ammonium disinfectants were all positive in the 16 strains of MRSA ,and positive rates of mecA ,aac（6′）/aph（2″） ,aph（3′）‐Ⅲ ,ermB ,tetM were all 100% ,but resistance genes to mupirocin were all negative .Moreover ,positive mode （mecA＋ aac（6′）/aph（2″）＋ aph（3′）‐Ⅲ ＋ ermB＋tetM＋ qacA） was positive in 13 strains（81 .3% ） .In addition ,the sample cluster analysis suggested that 13 strains （strain No .1 ,2 ,3 ,4 ,6 ,7 ,10 ,11 ,12 ,13 ,14 ,15 ,16） were the same clone ,and strains No .8 and 9 were the other clone .Relationship of 16 strains of MRSA and MRSA‐ref was close ,but relationship of 16 strains of MRSA and MSSA‐ref was far .CONCLUSION Pathogenicity of this group of MRSA carrying adhesion ,leucocidin ,andα‐hemolysin was strong .Resistance rates of this group of MRSA to 10 kinds of antimicrobials were all 100% ,which was identical to high positive rates of resistance genes .However ,mupirocin remained inhibitory to this group of M RS A .

关 键 词：耐甲氧西林金黄色葡萄球菌 黏附素 杀白细胞素 α溶血素 耐药基因 样本聚类分析 

分 类 号：R378[医药卫生—病原生物学]