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[2010 - 2014年间在某大学医院分离的肠杆菌科菌株的碳青霉烯酶基因及分子流行病学调查]

[Investigation of carbapenemase genes and molecular epidemiology of Enterobacteriaceae strains isolated between 2010-2014 in a university hospitals].

作者信息

Kutlu Hüseyin Haydar, Us Ebru, Tekeli Alper

机构信息

Usak University Faculty of Medicine, Department of Medical Microbiology, Usak, Turkey.

Ankara University Faculty of Medicine, Department of Medical Microbiology, Ankara, Turkey.

出版信息

Mikrobiyol Bul. 2018 Jan;52(1):1-12. doi: 10.5578/mb.66156.

Abstract

The worldwide spread of carbapenemase producing Enterobacteriaceae isolates has become a major threat of public health. This worrisome situation leads the development of new methods for carbapenemase screening, detection, prevention of spread and epidemiological data collection as mandatory. In this study, it was aimed to investigate existence and distribution of carbapenemase-encoding genes (CEGs) among carbapenem-resistant Enterobacteriaceae isolated from various clinical samples in Ankara University Faculty of Medicine, Ibni Sina Hospital, Central Microbiology Laboratory between June 2010-May 2014 and detect their clonal relationship. A total of 112 non-repetitive Enterobacteriaceae isolates which were intermediate or resistant to ertapenem were identified by using Phoenix (BD Diagnostic Systems, Sparks, USA) automated microbiology system. After DNA extraction from the isolates, 11 carbapenemase-encoding genes (CEGs) (bla, bla, bla, bla, bla, bla, bla, bla, bla bla ve bla) were detected with PCR. The clonal relationship among the isolates was determined by PFGE method following digestion with Xbal DNA macrorestriction endonuclease. Among 112 isolates Klebsiella pneumoniae was the most frequent (n= 79, 70.5%) bacteria followed by Escherichia coli (n= 15, 13.4%), Enterobacter cloacae (n= 10, 8.9%), Enterobacter aerogenes (n= 4, 3.6%) and Klebsiella oxytoca (n= 4, 3.6%) respectively. bla was the most frequent gene detected. Among 83 (74.1%) isolates bla was detected alone and in 7 (6.3%) of the isolates it was identified with bla gene coexistence. bla gene was identified as the second most frequent CEG among the isolates. bla gene was detected positive in 9 (8%) isolates. bla gene was identified in 2 (1.8%) isolates. Ten of the K.pneumoniae isolates with identical PFGE pattern were named as pulsotype B. These isolates were found to be similar in terms of isolate location, isolation dates, antibiotic resistance patterns and the carbapenemase genes they carry, and are considered to be potential outbreak isolates originated from intensive care units. On the other hand CEGs were found in the clinical samples obtained from five out-patients suggesting that community-acquired infections may also arise due to carbapenemase producing Enterobacteriaceae in our country where blaOXA-48 producers are endemic. According to this study, bla producing gram negative bacteria were frequent in our hospital. The prevalance of bla gene among metallo-beta-lactamases and coexistence with bla gene was remarkable. The frequency of blaNDM producing isolates in our hospital was not detected as high yet. In this study, the identification of carbapenemase producing bacteria as outbreak strains in our hospital indicated that cross-sectional surveillance for carbapenemase-producing bacteria from each patient was valuable in terms of early diagnosis of outbreaks.

摘要

产碳青霉烯酶肠杆菌科细菌分离株在全球范围内的传播已成为公共卫生的重大威胁。这种令人担忧的情况使得开发新的碳青霉烯酶筛查、检测、传播预防和流行病学数据收集方法成为当务之急。本研究旨在调查2010年6月至2014年5月间在安卡拉大学医学院伊本·西那医院中央微生物实验室从各种临床样本中分离出的耐碳青霉烯肠杆菌科细菌中碳青霉烯酶编码基因(CEGs)的存在和分布情况,并检测它们的克隆关系。使用Phoenix(美国BD诊断系统公司,斯帕克斯)自动化微生物系统共鉴定出112株对厄他培南呈中介或耐药的非重复性肠杆菌科细菌分离株。从这些分离株中提取DNA后,通过PCR检测到11种碳青霉烯酶编码基因(CEGs)(bla、bla、bla、bla、bla、bla、bla、bla、bla、bla和bla)。在用Xbal DNA大片段限制性内切酶消化后,通过PFGE方法确定分离株之间的克隆关系。在112株分离株中,肺炎克雷伯菌是最常见的细菌(n = 79,70.5%),其次是大肠埃希菌(n = 15,13.4%)、阴沟肠杆菌(n = 10,8.9%)、产气肠杆菌(n = 4,3.6%)和产酸克雷伯菌(n = 4,3.6%)。检测到的最常见基因是bla。在83株(74.1%)分离株中单独检测到bla,在7株(6.3%)分离株中它与bla基因共存。bla基因被确定为分离株中第二常见的CEG。在9株(8%)分离株中检测到bla基因呈阳性。在2株(1.8%)分离株中鉴定出bla基因。10株具有相同PFGE模式的肺炎克雷伯菌分离株被命名为脉冲型B。这些分离株在分离部位、分离日期、抗生素耐药模式以及它们携带的碳青霉烯酶基因方面相似,被认为是源自重症监护病房的潜在暴发分离株。另一方面,在从5名门诊患者获得的临床样本中发现了CEGs,这表明在我国blaOXA - 48产生菌为地方流行的情况下,产碳青霉烯酶肠杆菌科细菌也可能引起社区获得性感染。根据本研究,我院产bla的革兰阴性菌很常见。bla基因在金属β - 内酰胺酶中的流行率以及与bla基因的共存情况很显著。我院产blaNDM分离株的频率尚未检测到很高。在本研究中,将我院产碳青霉烯酶细菌鉴定为暴发菌株表明,对每位患者的产碳青霉烯酶细菌进行横断面监测对于暴发的早期诊断很有价值。

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