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[通过等电聚焦和聚合酶链反应对社区获得性尿源大肠杆菌分离株中ESBL类型的研究]

[Investigation of ESBL types in community acquired urinary Escherichia coli isolates by isoelectric focusing and polymerase chain reaction].

作者信息

Dağlar Duygu, Ongüt Gözde, Oğünç Dilara, Ozhak Baysan Betil, Demirbakan Hadiye, Sepin Özen Nevgün, Gültekin Meral

机构信息

Akdeniz Üniversitesi Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, Antalya, Türkiye.

出版信息

Mikrobiyol Bul. 2010 Jul;44(3):367-74.

Abstract

The aim of this study was to determine the extended-spectrum beta-lactamase (ESBL) types by isoelectric focusing (İEF) and polymerase chain reaction (PCR) methods in 56 Escherichia coli strains isolated from urine samples of patients with community-acquired urinary tract infection and determined as ESBL positive with the phenotypic screening tests (E test and combined disk method). IEF revealed that most of the strains produced 1 to 3 different bands, mostly at the isoelectric points 8.2 (n= 44, 79%) compatible with CTX-M. Twenty four (43%) isolates had CTX-M and TEM enzyme bands together, 16 (29%) isolates had only CTX-M enzyme bands, 3 (5%) isolates had CTX-M, TEM, SHV bands, one had CTX-M and SHV enzyme bands together, and one had only TEM band. Eleven E.coli strains did not yield any enzyme bands. PCR analysis revealed that 93% (n= 52) of the isolates had CTX-M, 64% (n= 36) had TEM and 11% (n= 6) had SHV, while 29 (52%) had CTX-M + TEM, three had CTX-M + SHV, and three had CTX-M + TEM + SHV genes together. PER-1 type beta-lactamases were not detected by PCR method. PCR analysis of the eleven strains that yielded no band in IEF showed that 5 strains had CTX-M + TEM, 3 had CTX-M and 3 had TEM enzyme genes. The consistency between IEF and PCR methods for the determination of CTX-M, TEM and SHV enzymes was 85%, 78% and 67%, respectively. Genes encoding ESBL's are usually located on transferrable plasmids that may also carry other resistance determinants. Thus detection of beta-lactamase enzyme types in ESBL positive bacteria is important for the choice of appropriate antimicrobial agents for treatment.

摘要

本研究的目的是通过等电聚焦(IEF)和聚合酶链反应(PCR)方法,对从社区获得性尿路感染患者尿液样本中分离出的56株大肠埃希菌进行超广谱β-内酰胺酶(ESBL)类型测定,这些菌株经表型筛选试验(E试验和复合纸片法)确定为ESBL阳性。IEF显示,大多数菌株产生1至3条不同的条带,主要在与CTX-M兼容的等电点8.2处(n = 44,79%)。24株(43%)分离株同时具有CTX-M和TEM酶条带,16株(29%)分离株仅有CTX-M酶条带,3株(5%)分离株具有CTX-M、TEM、SHV条带,1株同时具有CTX-M和SHV酶条带,1株仅有TEM条带。11株大肠埃希菌未产生任何酶条带。PCR分析显示,93%(n = 52)的分离株具有CTX-M,64%(n = 36)具有TEM,11%(n = 6)具有SHV,而29株(52%)同时具有CTX-M + TEM,3株具有CTX-M + SHV,3株同时具有CTX-M + TEM + SHV基因。PCR方法未检测到PER-1型β-内酰胺酶。对在IEF中未产生条带的11株菌株进行PCR分析显示,5株具有CTX-M + TEM,3株具有CTX-M,3株具有TEM酶基因。IEF和PCR方法在测定CTX-M、TEM和SHV酶方面的一致性分别为85%、78%和67%。编码ESBL的基因通常位于可转移质粒上,这些质粒也可能携带其他耐药决定子。因此,检测ESBL阳性细菌中的β-内酰胺酶类型对于选择合适的抗菌药物进行治疗很重要。

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