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[圣地亚哥德孔波斯特拉医疗保健区域中产氨苄西林酶奇异变形杆菌的质粒介导:rep-PCR和基质辅助激光解吸电离飞行时间质谱的分子及流行病学分析]

[Plasmid-mediated AMPc producing Proteus mirabilis in the Health Care Area of Santiago de Compostela: molecular and epidemiological analysis by rep-PCR and MALDI-TOF].

作者信息

Treviño Mercedes, Navarro Daniel, Barbeito Gema, Areses Paloma, García-Riestra Carlos, Regueiro Benito J

机构信息

Servicio de Microbiología, Complejo Hospitalario Universitario de Santiago de Compostela, Spain.

出版信息

Rev Esp Quimioter. 2012 Jun;25(2):122-8.

Abstract

INTRODUCTION

Proteus mirabilis is an important pathogen isolated from both community-acquired and health-care associated infections. Acquired AmpC-type beta-lactamases represent an important mechanism of resistance to extended-spectrum cephalosporins and are emerging in several European countries. The objective of this work was to know the prevalence of acquired AmpC beta-lactamase producing P. mirabilis over the last three years and eight months and their clonal relationships comparing MALDI-TOF and automated rep-PCR results.

METHODS

P. mirabilis isolates (n= 1,396) were obtained from routine cultures at the University Hospital Complex of Santiago de Compostela from January 2006 to August 2009. Identification to the species level and antimicrobial susceptibility testing were achieved with Vitek 2. The isolates showing intermediate or total resistance to amoxicillin-clavulanic and cefoxitin, cefotaxime or ceftazidime were selected for AmpC phenotypic detection by double-disk synergy test, and molecular confirmation by multiplex PCR. Molecular typing of the isolates was performed by automated rep-PCR and MALDI-TOF.

RESULTS

For the last three years and eight months, the prevalence of AmpC-producing P. mirabilis increased from 0.17% to 4.5%, mainly associated with urinary tract infection in elderly outpatients. In all cases, plasmidic AmpC belonging to LAT/CMY lineage were detected. A high genetic variability was seen with both, rep-PCR and MALDI-TOF MS.

CONCLUSIONS

AmpC-producing P. mirabilis is an emergent pathogen. The high genetic variability detected suggests that the spread of the resistance mechanism is more probable than a clone dispersion. Automated rep-PCR and MALDI-TOF MS show as fast and decisive methods for bacterial strain typing in clinical microbiology laboratories.

摘要

引言

奇异变形杆菌是一种从社区获得性感染和医疗保健相关感染中分离出的重要病原体。获得性AmpC型β-内酰胺酶是对广谱头孢菌素耐药的重要机制,并且在几个欧洲国家中不断出现。这项工作的目的是了解过去三年零八个月中产生获得性AmpCβ-内酰胺酶的奇异变形杆菌的流行情况,以及比较基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)和自动重复聚合酶链反应(rep-PCR)结果的克隆关系。

方法

从2006年1月至2009年8月在圣地亚哥德孔波斯特拉大学医院综合大楼的常规培养物中获得奇异变形杆菌分离株(n = 1396)。使用Vitek 2进行菌种鉴定和抗菌药物敏感性测试。通过双纸片协同试验选择对阿莫西林-克拉维酸、头孢西丁、头孢噻肟或头孢他啶表现出中度或完全耐药的分离株进行AmpC表型检测,并通过多重聚合酶链反应进行分子确认。通过自动rep-PCR和MALDI-TOF对分离株进行分子分型。

结果

在过去三年零八个月中,产AmpC的奇异变形杆菌的流行率从0.17%增加到4.5%,主要与老年门诊患者的尿路感染有关。在所有病例中,均检测到属于LAT/CMY谱系的质粒AmpC。rep-PCR和MALDI-TOF MS均显示出高度的遗传变异性。

结论

产AmpC的奇异变形杆菌是一种新兴病原体。检测到的高遗传变异性表明耐药机制的传播比克隆传播更有可能。自动rep-PCR和MALDI-TOF MS显示为临床微生物学实验室中细菌菌株分型的快速且决定性的方法。

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