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目的分析一所三级教学医院临床分离的20株多药耐药铜绿假单胞菌(MDR-PAE)的菌株亲缘关系。方法收集2009年2月-2010年4月,从医院住院患者分离出的MDR-PAE 20株,采用聚合酶链反应(PCR)及序列分析方法对42种β-内酰胺酶基因、氨基糖苷类修饰酶基因、甲基化酶基因、可移动遗传元件遗传标记以及毒力因子基因进行检测,并将检测结果作聚类分析以探讨MDR-PAE的菌株亲缘关系。结果 20株MDR-PAE中,TEM、PER、DHA、OXA-10群等4种β-内酰胺酶基因的阳性检出率分别为25.0%、20.0%、5.0%及20.0%,膜孔蛋白编码基因oprD2缺失率为100.0%;tnp513/ISCR1I、S26I、Spa7及intⅠ1等4种转座子、插入序列和整合子遗传标记基因的阳性检出率分别为20.0%、20.0%、30.0%及30.0%;氨基糖苷类修饰酶基因aac(3)-Ⅱ、aac(6′)-Ⅰb、aac(6′)-Ⅱ、ant(3″)-Ⅰ及ant(2″)-Ⅰ的阳性检出率分别为40.0%、10.0%、20.0%、15.0%及10.0%,甲基化酶基因rmtB的阳性率为15.0%;以42种β-内酰胺酶基因、氨基糖苷类修饰酶基因、甲基化酶基因和可移动遗传元件遗传标记以及毒力因子基因作检测结果的样本聚类分析显示,编号为No.2、8等2株,4、5、6等3株及1、10、11、12、14、15、20等7株为三群克隆传播。结论多药耐药铜绿假单胞菌株在该院不同科室间存在3群克隆传播株,加强对MDR-PAE耐药基因检测并对结果进行聚类分析对其医院感染的实时监测很有意义。
Objective To analyze the genetic relationship of 20 multidrug-resistant Pseudomonas aeruginosa (MDR-PAE) isolates clinically isolated in a tertiary teaching hospital. Methods Twenty MDR-PAE isolates were collected from inpatients in our hospital from February 2009 to April 2010. PCR-based polymerase chain reaction (PCR) and sequence analysis were used to analyze the polymorphisms of 42 β-lactamase genes, aminoglycosides Modified enzyme gene, methylase gene, genetic markers of mobile genetic elements and virulence factor gene were detected, and the test results for cluster analysis to explore the MDR-PAE strain genetic relationship. Results The positive detection rates of four β-lactamase genes such as TEM, PER, DHA and OXA-10 in 20 MDR-PAEs were 25.0%, 20.0%, 5.0% and 20.0%, respectively. The deletion rate of oprD2 was 100.0%. The positive rates of insertions and integron genetic markers were 20.0%, 20.0%, 30.0% and 30.0% respectively for 4 transposons including tnp513 / ISCR1I, S26I, Spa7 and intⅠ1, ; The positive detection rate of aminoglycoside modified enzyme genes aac (3) -Ⅱ, aac (6 ’) - Ⅰb, aac (6’) - Ⅱ, ant (3 ") - Ⅰ and ant Respectively. The positive rates of methylated enzyme gene rmtB were 15.0%, 40.0%, 10.0%, 20.0%, 15.0% and 10.0%, respectively. The positive rates of 42 β-lactamase genes, aminoglycoside- Cluster analysis of the results of genetic markers of the enzyme gene and the movable genetic element and the virulence factor gene showed that 2 strains of No.2 and 8 strains, 3 strains of 4, 5, 6 and 1 strains of 10, 10, Seven, 12, 14, 15, 20 and other seven clones spread. Conclusions There are three groups of cloned multidrug-resistant Pseudomonas aeruginosa strains in different departments of the hospital. To strengthen the detection of multidrug-resistant Pseudomonas aeruginosa strains and to cluster MDR-PAE resistance genes in real-time monitoring of nosocomial infections is of great significance.