耐碳青霉烯类肺炎克雷伯菌的耐药机制及同源性分析  

Resistance mechanisms and homology analysis in carbapenem-resistant Klebsiella pneumoniae

作　　者：黄亚雨 涂海健 郑磊[3] HUANG Yayu;TU Haijian;ZHENG Lei(Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学,广州510515 [2]南方医科大学附属莆田医院 [3]南方医科大学南方医院

出　　处：《山东医药》2018年第40期28-31,共4页Shandong Medical Journal

基　　金：福建省自然科学基金项目(2015J0105)

摘　　要：目的分析耐碳青霉烯类肺炎克雷伯菌(CRKP)的耐药机制及其同源性,为临床治疗此类耐药菌株感染提供依据。方法收集33株临床分离的CRKP菌株,采用法国梅里埃VITEK 2-compact分析仪进行细菌检测和药敏试验,改良Hodge试验和EDTA纸片协同试验检测耐药表型,PCR法扩增检测碳青霉烯酶基因(8种)及膜孔蛋白基因,ERIC-PCR法及多位点序列分型(MLST)法进行同源性分析及基因分子分型。结果 33株CRKP对米诺环素、复方新诺明、丁胺卡那霉素、庆大霉素、妥布霉素、舒普森、呋喃妥因、氨苄西林、哌拉西林/他唑巴坦、环丙沙星、左旋氧氟沙星、氨苄西林钠舒巴坦钠、头孢曲松、头孢唑林、头孢吡肟和头孢他啶16种抗菌药物存在多重耐药。改良Hodge试验发现,产金属β-内酰胺酶菌株30株,EDTA纸片协同试验发现产金属β-内酰胺酶菌株1株。PCR检测结果显示,携带KPC基因32株,携带SHV基因30株,携带NDM-1基因1株,未检测出其他耐药基因; ompk35膜孔蛋白基因缺失1株,ompk36膜孔蛋白基因缺失3株。ERIC-PCR法及MLST法结果显示,ST11/A型29株,ST15/B型2株,ST11/C和ST2193/D各1株。结论耐碳青霉烯类肺炎克雷伯菌耐药机制与其携带KPC耐药基因并产生KPC酶有关;大部分CRKP为ST11/A型,同源性程度高,可在院内克隆性传播。Objective To study the carbapenem resistance mechanism and the homology of carbapenem-resistant Klebsiella pneumoniae(CRKP)in order to provide corresponding infection control measures.Methods Clinical isolated strains of CRKP were collected.The bacterial identification and antimicrobial susceptibility testing were carried out by automatic microbiological analyzer-Merie VITEK from France.The resistance phenotypes in Klebsiella pneumoniae were detected by modified hodge tests and ethylenediamine tetraacetic acid(EDTA)disk synergy tests.Eight carbapenemase genes and membranoprotein genes were detected by PCR.The homology analysis and genotypes of the strains were individually analyzed by multilocus sequence typing(MLST)and enterbacterial repetitive intergenic consensus-PCR(ERIC-PCR).Results Thirty-three strains of CRKP were collected.All strains had multiple resistance to 16 antibiotics,including Minocycline,compound ciprofloxacin,amikacin,gentamicin,tobramycin,sampson,furantoin,ampicillin,piperacillin/azobactin,ciprofloxacin,levofloxacin,ampicillin sodium and sulbactum sodium,ceftriaxone,cefazolin,cefepime and cefazididin;thirty strains were tested positive by modified hodge test and one strain was positive by EDTA-disk synergy test.Thirty-two isolates carried KPC genes,and thirty isolates carried SHV genes.NDM-1 gene was detected in one isolate.One strain did not produce Ompk35 membranoprotein,and three strains were negative with Ompk36 membranoprotein gen.In addition,all 33 isolates did not produced VIM,IMP,OXA-48,GES,and GIM.MLST and ERIC-PCR electrophores results showed that ST11/A type was found in 29 strains,ST15/B type in 2,ST11/C type in 1,and ST2193/D type in 1.Conclusion The strains of CRKP carry KPC resistance genes which produce KPC enzymes,and most of the strains of CRKP are classified into ST11/A with high homology,and could be clonally transmitted inside the hospital.

关 键 词：肺炎克雷伯菌 碳青霉烯酶 KPC耐药基因 KPC酶耐药性 同源性 

分 类 号：R378[医药卫生—病原生物学]