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从土耳其一家三级医院的携带者和临床样本中分离出的耐万古霉素粪肠球菌菌株的分子特征

Molecular characterization of vancomycin-resistant Enterococcus faecium strains isolated from carriage and clinical samples in a tertiary hospital, Turkey.

作者信息

Gozalan Aysegul, Coskun-Ari Fatma Filiz, Ozdem Birsen, Unaldi Ozlem, Celikbilek Nevreste, Kirca Fisun, Aydogan Sibel, Muderris Tuba, Guven Tumer, Acikgoz Ziya Cibali, Durmaz Riza

机构信息

Atatürk Education and Research Hospital, Medical Microbiology Laboratory, Ankara, Turkey.

National Molecular Microbiology Reference Laboratory, Public Health Institution of Turkey, Ankara, Turkey.

出版信息

J Med Microbiol. 2015 Jul;64(7):759-766. doi: 10.1099/jmm.0.000088. Epub 2015 May 14.

Abstract

This study aimed to determine the presence of vancomycin resistance (vanA and vanB) and virulence genes (esp, asa1, gelE, ace, hyl, cylA, cpd and ebpA) in vancomycin-resistant Enterococcus faecium (VREfm) strains and to analyse the clonal relationships among the strains. E. faecium strains were identified from rectal and clinical specimens by biochemical tests and the API-20 Strep kit. Susceptibility testing was performed using disc-diffusion and broth-dilution methods. PFGE was used for molecular typing of the VREfm strains. The vancomycin resistance and virulence genes were amplified by two-step multiplex PCR. All 55 VREfm isolates were resistant to penicillin G, ampicillin and high-level gentamicin but were susceptible to quinupristin/dalfopristin and linezolid. Multiplex PCR analysis indicated that all isolates harboured vanA and that 41 (75 %) were positive for virulence genes. The esp gene was the most common virulence factor and was detected in nine (41 %) invasive and 32 (96.7 %) non-invasive isolates. Multiple virulence genes were observed only in two non-invasive isolates; one harboured esp and ebpA and the other harboured esp, ebpA, asa1, gelE and cpd. PFGE typing yielded 16 different types, seven of which were clusters with two to 14 strains each. The clustering rates of the rectal swab, blood and urine isolates were 72.7 %, 61.5 % and 87.5 %, respectively. The genetic similarity observed among the VREfm isolates indicated cross-transmission in the hospital. Further studies on the virulence factors present in the strains might provide insight into the acquisition of these traits and their contribution to increased prevalence of VREfm.

摘要

本研究旨在确定耐万古霉素屎肠球菌(VREfm)菌株中万古霉素耐药基因(vanA和vanB)及毒力基因(esp、asa1、gelE、ace、hyl、cylA、cpd和ebpA)的存在情况,并分析这些菌株之间的克隆关系。通过生化试验和API-20 Strep试剂盒从直肠和临床标本中鉴定屎肠球菌菌株。采用纸片扩散法和肉汤稀释法进行药敏试验。PFGE用于VREfm菌株的分子分型。通过两步多重PCR扩增万古霉素耐药基因和毒力基因。所有55株VREfm分离株均对青霉素G、氨苄西林和高水平庆大霉素耐药,但对奎奴普丁/达福普汀和利奈唑胺敏感。多重PCR分析表明,所有分离株均携带vanA,41株(75%)毒力基因呈阳性。esp基因是最常见的毒力因子,在9株(41%)侵袭性分离株和32株(96.7%)非侵袭性分离株中被检测到。仅在2株非侵袭性分离株中观察到多个毒力基因;一株携带esp和ebpA,另一株携带esp、ebpA、asa1、gelE和cpd。PFGE分型产生了16种不同类型,其中7种为聚类,每个聚类有2至14株菌株。直肠拭子、血液和尿液分离株的聚类率分别为72.7%、61.5%和87.5%。VREfm分离株之间观察到的遗传相似性表明在医院内存在交叉传播。对菌株中存在的毒力因子进行进一步研究可能有助于深入了解这些性状的获得及其对VREfm患病率增加的作用。

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