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目的探讨耐碳青霉烯类肺炎克雷伯菌耐药基因及可移动遗传元件遗传标记的携带情况及菌株之间的亲缘关系。方法收集2013年1-12月医院住院患者中分离到的20株耐碳青霉烯类肺炎克雷伯菌,用gyrA与parC基因测序确认菌种,再采用聚合酶链反应(PCR)及序列分析的方法分析56种获得性耐药基因、12种MGEs标记、膜孔蛋白基因(ompK35、ompK36)以及喹诺酮类作用靶位基因(gyrA、parC),最后对检测结果作样本聚类分析。结果 20株耐碳青霉烯类肺炎克雷伯菌共检出6种β-内酰胺类药物获得性耐药基因、2种膜孔蛋白基因、5种氨基糖苷类药物获得性耐药基因、2种喹诺酮类药物作用靶位基因、1种喹诺酮类药物获得性耐药基因、8种可移动遗传元件标记;样本聚类分析提示,10号为孤立株,其余19株呈明显的聚集性(A家族);A家族可分为A1和A2;A1又可分为A1-1和A1-2;A1-1再可分为A1-1-1和A1-1-2小家族;A1-1-1和A1-1-2小家族聚集性更为明显,并分别出现克隆播散。结论获得性耐药基因与管家基因检测结果提供了本组菌株耐药的遗传基础,并且与耐药表型相对应,样本聚类分析是监控医院感染实用手段。
Objective To investigate the resistance of carbapenem-resistant Klebsiella pneumoniae and the genetic markers of portable genetic elements and the genetic relationship between strains. Methods Twenty isolates of Klebsiella pneumoniae resistant to carbapenem were collected from inpatients from January to December in 2013. The strains were identified by gyrA and parC gene sequencing and then sequenced by polymerase chain reaction (PCR) and sequencing Analysis of 56 kinds of acquired resistance genes, 12 kinds of MGEs markers, membrane porin gene (ompK35, ompK36) and quinolones role target gene (gyrA, parC), and finally the test results for sample clustering analysis. Results 20 strains of carbapenem-resistant Klebsiella pneumoniae were detected six acquired β-lactam drug resistance genes, two kinds of membrane pore protein genes, five aminoglycoside-acquired resistance genes, Two kinds of quinolone target genes and one quinolone resistance gene were obtained, and eight kinds of mobile genetic elements were labeled. Cluster analysis indicated that isolate 10 was isolated and the remaining 19 were obviously aggregated A family); A family can be divided into A1 and A2; A1 can be divided into A1-1 and A1-2; A1-1 can be divided into A1-1-1 and A1-1-2 small family; A1-1 -1 and A1-1-2 small familial aggregation is more obvious, and clonal dissemination appeared. Conclusion The acquired resistance gene and housekeeping gene test results provide the genetic basis of drug resistance in this group of patients. Corresponding to the resistant phenotype, sample cluster analysis is a practical means of monitoring hospital infection.