宁夏地区多重耐药铜绿假单胞菌的临床耐药分布及耐药研究
发布时间：2018-09-06 07:12
【摘要】：目的研究铜绿假单胞菌的临床分布和抗菌药物的耐药性,从分子水平探讨其产生多重耐药的可能机制和感染的同源性,从而为有效地预防和控制多重耐药铜绿假单胞菌感染提供科学的参考依据。方法1.收集并保存宁夏医科大学附属医院自2011年9月至2012年9月期间临床分离的非重复PA菌株285株,采用法国梅里埃公司全自动微生物分析仪(VITEK 2 COMPACT)鉴定细菌;采用K-B法检测PA临床分离株对14种常用抗生素的药物敏感试验,应用WHONET5.6软件对上述药物敏感结果进行统计分析,筛选出多重耐药铜绿假单胞菌。2.将筛选出的124株MDRP菌株通过外排泵抑制剂利血平对羧苄西林、红霉素、亚胺培南及庆大霉素的琼脂稀释法抑制试验分别检测铜绿假单胞菌的Mex AB-Opr M、Mex CD-Opr J、Mex EF-Opr N、Mex XY-Opr M四种外排泵的表型,同时检测加入利血平后四种药物对铜绿假单胞菌MIC值的变化;应用聚合酶链反应(PCR)技术检测外排泵基因。3.将筛选出的124株MDRP菌株,通过PCR检测整合酶基因,将IntⅠ检测阳性菌株继续采用PCR方法扩增可变区;采用PCR-RFLP分析并确定菌株可变区类型。4.应用脉冲场凝胶电泳(PFGE)技术对多重耐药铜绿假单胞菌进行同源性分析。结果1.2011年9月至2012年9月我院共分离出285株铜绿假单胞菌,标本的来源主要是痰液(65.5%),科室的分布主要集中在ICU(34.7%),耐药结果显示:临床分离的铜绿假单胞菌对多粘菌素B敏感,对亚胺培南、头孢他啶等耐药率低,在临床治疗中可以选择联合用药;筛选出的124株MDRP亦主要来自痰液(62.9%),主要分布在ICU(22.6%)。2.外排泵表型阳性结果为:Mex AB-Opr M74株、Mex XY-Opr M28株、Mex EF-Opr N12株和Mex CD-Opr J10株;124株MDRP菌株中74株Mex AB-Opr M外排泵表型阳性。3.检测124株MDRP其中Ⅰ类整合子阳性菌株有87株,未检测出Ⅱ类整合子;87株IntⅠ基因阳性的菌株中有53株有效的扩增出Ⅰ类整合子可变区,通过测序发现6种形式的耐药基因盒。4.对124株MDRP进行脉冲场凝胶电泳(PFGE)分析得到8个基因型,其中主要的有5个基因型。结论1.在临床常用抗生素中,铜绿假单胞菌临床分离株对多粘菌素B最敏感,对复方新诺明的耐药率最高。标本来源主要是痰液,PA主要分布在ICU。2.四种外排泵表型中最常见的是Mex AB-Opr M,推测其可能是铜绿假单胞菌多重耐药的重要机制。3.多重耐药铜绿假单胞菌整合子的分布类型以Ⅰ类整合子为主,整合子中的氨基糖甙类修饰酶基因和β-内酰胺类基因的存在与多药耐药铜绿假单胞菌的多重耐药密切相关。4.在基因水平上通过PFGE对MDRP进行分析,得出MDRP感染来源、传播途径及分布规律,对感染流行的监测提供可靠依据。
[Abstract]:Objective to study the clinical distribution and antimicrobial resistance of Pseudomonas aeruginosa and to explore the possible mechanism of multidrug resistance and the homology of infection at molecular level. It provides a scientific reference for the prevention and control of multidrug resistant Pseudomonas aeruginosa infection. Method 1. From September 2011 to September 2012, 285 strains of non-repetitive PA strains were collected and preserved in the affiliated Hospital of Ningxia Medical University. The bacteria were identified by VITEK 2 COMPACT. K-B method was used to detect the drug sensitivity of clinical isolates of PA to 14 common antibiotics. The results of drug sensitivity were statistically analyzed by WHONET5.6 software, and the multidrug resistant Pseudomonas aeruginosa was screened out. Carbenicillin and erythromycin were separated from 124 strains of MDRP by reserpine, an efflux pump inhibitor. Imipenem and gentamicin Agar dilution inhibition tests were used to detect the phenotypes of four efflux pumps of Pseudomonas aeruginosa (Mex AB-Opr MMex CD-Opr JnMex EF-Opr NMex XY-Opr M), and the changes of MIC value of four drugs on Pseudomonas aeruginosa after adding reserpine. Polymerase chain reaction (PCR) technique was used to detect the efflux pump gene. 124 strains of MDRP were screened, the integrase gene was detected by PCR, the Int 鈪，
本文编号：2225629
[Abstract]:Objective to study the clinical distribution and antimicrobial resistance of Pseudomonas aeruginosa and to explore the possible mechanism of multidrug resistance and the homology of infection at molecular level. It provides a scientific reference for the prevention and control of multidrug resistant Pseudomonas aeruginosa infection. Method 1. From September 2011 to September 2012, 285 strains of non-repetitive PA strains were collected and preserved in the affiliated Hospital of Ningxia Medical University. The bacteria were identified by VITEK 2 COMPACT. K-B method was used to detect the drug sensitivity of clinical isolates of PA to 14 common antibiotics. The results of drug sensitivity were statistically analyzed by WHONET5.6 software, and the multidrug resistant Pseudomonas aeruginosa was screened out. Carbenicillin and erythromycin were separated from 124 strains of MDRP by reserpine, an efflux pump inhibitor. Imipenem and gentamicin Agar dilution inhibition tests were used to detect the phenotypes of four efflux pumps of Pseudomonas aeruginosa (Mex AB-Opr MMex CD-Opr JnMex EF-Opr NMex XY-Opr M), and the changes of MIC value of four drugs on Pseudomonas aeruginosa after adding reserpine. Polymerase chain reaction (PCR) technique was used to detect the efflux pump gene. 124 strains of MDRP were screened, the integrase gene was detected by PCR, the Int 鈪，
本文编号：2225629
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