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目的了解耐碳青霉烯类肺炎克雷伯菌(CRKP)对β-内酰胺类与氨基糖苷类药物的耐药相关基因与可移动遗传元件存在状况,以及获得性耐药相关基因与可移动遗传元件存在的相互关系。方法收集2014年1-12月住院患者标本中分离到的20株CRKP,用gyrA测序后BLASTn比对确认菌种,再采用聚合酶链反应(PCR)法分析40种β-内酰胺酶基因、6种氨基糖苷类修饰酶、6种16SrRNA甲基化酶、12种可移动遗传元件遗传标记和blaKPC型与ISKpn6连锁检测,并对检测结果作指标聚类分析。结果 20株CRKP对9种β-内酰胺类与氨基糖苷类均耐药;获得性耐药相关基因与可移动遗传元件标记每株均有阳性发现,共检出4种β-内酰胺酶基因(blaTEM、blaSHV、blaKPC、blaIMP),1种氨基糖苷类修饰酶基因(aac(3)-Ⅱ)、1种16SrRNA甲基化酶基因(rmtB),8种可移动遗传元件遗传标记(traA、trbC、tnp513、IS26、IS903、ISEcp1、ISKpn6、intⅠ1);5株CRKP blaKPC-ISKpn6连锁检测为阳性;指标聚类分析提示,β-内酰胺酶基因blaIMP与tnp513强关联;β-内酰胺酶基因blaKPC、blaSHV及16S rRNA甲基化酶基因rmtB与ISKpn6、ISEcp1、traA强关联;氨基糖苷类修饰酶基因aac(3)-Ⅱ与IS26、IS903、intⅠ1、trbC强关联。结论肺炎克雷伯菌耐药表型与基因型结果相符,对β-内酰胺类与氨基糖苷类药物的耐药与可移动遗传元件介导的blaTEM、blaSHV、blaKPC、blaIMP、aac(3)-Ⅱ、rmtB相关。
Objective To understand the existence of resistance-related genes and mobile genetic elements of carbapenem-resistant Klebsiella (CRKP) to β-lactams and aminoglycosides and the relationship between acquired resistance-related genes The relationship between the existence of genetic elements. Methods Twenty isolates of CRKP from inpatients from January to December 2014 were collected. The strains were identified by BLASTn sequencing after gyrA sequencing, and 40 kinds of β-lactamase genes were analyzed by polymerase chain reaction (PCR) Six kinds of aminoglycoside modified enzymes, six kinds of 16SrRNA methylase, genetic markers of 12 kinds of movable genetic elements, and the detection of blaKPC type and ISKpn6. The clustering analysis was made on the test results. Results Twenty CRKP strains were resistant to 9 β-lactams and aminoglycosides. The results showed that there were four positive β-lactamase genes (blaTEM, blaSHV, blaKPC and blaIMP), one aminoglycoside modified enzyme gene (aac (3) -II), one 16S rRNA methylase gene (rmtB), eight movable genetic elements (traA, The results of cluster analysis indicated that the blaIMP gene of β-lactamase was strongly associated with tnp513, the gene of β-lactamase was significantly higher than that of tnp513 (P <0.05) blaKPC, blaSHV and 16S rRNA methylase gene rmtB were strongly associated with ISKpn6, ISEcp1 and traA. The aminoglycoside-modifying enzyme gene aac (3) -II was strongly associated with IS26, IS903, intⅠ1 and trbC. Conclusion The resistant phenotypes of Klebsiella pneumoniae are consistent with those of genotypes. The resistance of blakes, blaSHV, blaKPC, blaIMP and aac (3) mediated by resistance and transmissible genetic elements of β-lactams and aminoglycosides, -Ⅱ, rmtB related.