耐碳青酶烯类肺炎克雷伯菌产酶耐药机制及同源性分析

发布时间：2018-07-14 10:39

【摘要】：目的：筛选重庆某三级医院临床分离耐碳青霉烯类肺炎克雷伯菌产酶耐药菌株,并对产酶机制及耐药菌株同源性进行分析,为临床治疗和控制耐碳青霉烯类肺炎克雷伯菌(CRKP)提供理论依据。方法：收集重庆某三级医院2003年-2013年临床分离肺炎克雷伯菌200株。用PCR的方法通过扩增khe基因对菌株进行鉴定；常规药敏纸片试验及MIC值测定筛选出CRKP;用改良Hodge试验和改良Carba NP试验筛选出产碳青霉烯酶菌株；PCR扩增耐药基因并对PCR产物进行测序对比；质粒结合实验研究细菌耐药基因的传播方式；用MLST和PFGE方法对细菌进行同源性分析。结果：从临床收集的肺炎克雷伯菌中共筛选出22株CRKP,大部分对厄他培南不敏感。22株CRKP中有6株产酶表型阳性且产B类酶,常见的产酶耐药基因扩增结果显示,6株携带耐药基因,经测序和网上比对结果显示,其中1株产酶菌株携带NDM-1基因,1株携带IMP-4基因,4株携带IMP-8基因。6株产酶菌株质粒结合试验均成功。MLST和PFGE对产酶菌株进行分子分型,6株CRKP表现出大小不一的同源性,6株产酶肺炎克雷伯菌一共分为4个基因型。结论：CRKP有多种耐药机制,产碳青霉烯酶是最常见的耐药机制之一。该医院产碳青霉烯酶耐药菌主要携带耐药基因IMP-8、IMP-4、OXA-1,并且耐药基因可以通过质粒进行传递。MLST和PFGE的结果显示此医院有主要的流行菌株,应引起临床各科室和医院感染控制部门的高度重视,避免发生医院感染的暴发流行。
[Abstract]:Objective: to screen the antibiotic resistant strains of Klebsiella pneumoniae from a three grade hospital in Chongqing, and to analyze the homology of the enzyme producing mechanism and resistant strains, and to provide a theoretical basis for the clinical treatment and control of Klebsiella pneumoniae (CRKP) in the clinical treatment and control of carbon Penicillium pneumonia (Klebsiella pneumoniae). 200 strains of Klebsiella pneumoniae were isolated from the bed. The strains were identified by Khe gene amplification by PCR method; CRKP was screened by conventional drug sensitive paper test and MIC value determination. The modified Hodge test and modified Carba NP test were used to screen out carbapenem producing strains; PCR amplification resistance gene was sequenced and the PCR products were sequenced; plasmid binding solid The transmission patterns of bacterial resistance genes were studied. The homology analysis of bacteria was carried out by MLST and PFGE methods. Results: 22 strains of CRKP were screened from Klebsiella pneumoniae collected clinically, and 6 of the.22 strains of unsensitive.22 strain CRKP were positive and produced B type enzymes, and 6 of the common enzyme resistant gene amplification results showed that 6 1 strains of enzyme producing strains carrying NDM-1 gene, 1 strains carrying IMP-4 gene and 4 strains of IMP-8 gene.6 producing enzyme strains were successfully carried out by.MLST and PFGE for molecular typing, and 6 strains of CRKP showed different homology, and 6 strains of klebber producing enzyme pneumonia. The bacteria are divided into 4 genotypes. Conclusion: CRKP has a variety of resistance mechanisms. Carbapenenecylenase is one of the most common mechanisms of resistance. Carbapenem resistant bacteria in the hospital mainly carry resistance genes IMP-8, IMP-4, OXA-1, and the resistance genes can transmit.MLST and PFGE through plasmids to show that the hospital has major epidemic bacteria. It should arouse the attention of clinical departments and hospital infection control departments to avoid the outbreak of nosocomial infection.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.5

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