Molecular diversity in mechanisms of carbapenem resistance in paediatric Enterobacteriaceae.
Molecular diversity in mechanisms of carbapenem resistance in paediatric Enterobacteriaceae.
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DOI：
10.1016/j.ijantimicag.2011.09.014
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发表时间：
2012-01
影响因子：
10.8
通讯作者：
Weissman SJ
中科院分区：
文献类型：
--
作者：
Little ML;Qin X;Zerr DM;Weissman SJ
肠杆菌科中碳青霉烯耐药性的发展影响了临床和实验室标准化研究所（CLSI）的指南、感染控制方法和治疗策略。儿科转诊中心碳青霉烯类耐药肠杆菌科（CRE）感染的临床、表型和基因型特征尚未得到充分描述。通过西雅图儿童医院（西雅图，WA）的临床微生物学实验室鉴定CRE。从病历中检索临床数据。对每种分离株进行耐药性检测、耐药决定簇聚合酶链反应（PCR）和大肠杆菌转化。使用多位点序列分型（MLST）和脉冲场凝胶电泳（PFGE）来表征菌株相关性。采用PCR扩增、测序及十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）等方法研究孔蛋白的变化。在2002年至2010年期间鉴定了6株CRE分离株。其耐药机制存在显著的分子多样性，包括质粒介导的丝氨酸碳青霉烯酶（KPC）和金属β-内酰胺酶（IMP）、染色体编码的β-内酰胺酶（SME）和超广谱β-内酰胺酶引起的孔蛋白改变。患者有潜在的健康状况，来自不同的地理区域。在一个案例中，系列分离株的PFGE记录了以前敏感菌株的耐药性发展。该菌株的分子研究鉴定了ompK 36基因中遗传移动的元件插入序列ISEcp 1的插入，如SDS-PAGE所示，赋予功能性孔蛋白改变。这是首次描述CTX-M-15阳性分离株中ISEcp 1对孔蛋白的破坏。这是迄今为止最大的儿科CRE报告。这种对人口统计学、表型和分子特征的不同描述突出了高危儿科患者中CRE感染的挑战，并且在儿科转诊中心关注新出现的耐药机制（包括膜改变）至关重要。
Development of carbapenem resistance in Enterobacteriaceae has impacted Clinical and Laboratory Standards Institute (CLSI) guidelines, infection control approaches and treatment strategies. The clinical, phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae (CRE) infections at paediatric referral centres are not well described. CRE were identified through the clinical microbiology laboratory at Seattle Children’s Hospital (Seattle, WA). Clinical data were retrieved from medical records. Resistance testing, polymerase chain reaction (PCR) for resistance determinants, and Escherichia coli transformation were carried out for each isolate. Multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used to characterise strain relatedness. PCR amplification and sequencing as well as sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) were used to investigate porin alterations. Six CRE isolates were identified between 2002 and 2010. Significant molecular diversity was documented in their mechanisms of resistance, including plasmid-mediated serine carbapenemase (KPC) and metallo-β-lactamase (IMP), chromosomally-encoded β-lactamase (SME) and porin alterations with extended-spectrum β-lactamases. Patients had underlying health conditions and were from geographically diverse regions. In one case, PFGE of serial isolates documented the development of resistance in a previously susceptible strain. Molecular investigation of this strain identified insertion of the genetic mobile element insertion sequence ISEcp1 in the ompK36 gene, conferring a functional porin alteration as demonstrated by SDS-PAGE. This is the first description of porin disruption by ISEcp1 in a CTX-M-15-positive isolate. This is the largest report of paediatric CRE to date. This diverse description of demographic, phenotypic and molecular characteristics highlights the challenge of CRE infections in high-risk paediatric patients and that attention to emerging resistance mechanisms (including membrane alteration) at paediatric referral centres is essential.
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作者：
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通讯作者：
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影响因子：
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作者：
Barlow, Miriam;Reik, Rebecca A.;Jacobs, Stephen D.;Medina, Monica;Meyer, Matthew P.;McGowan, John E., Jr.;Tenover, Fred C.
通讯作者：
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影响因子：
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通讯作者：
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