A single step multiplex PCR for identification of six diarrheagenic E. coli pathotypes and Salmonella
A single step multiplex PCR for identification of six diarrheagenic E. coli pathotypes and Salmonella
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DOI：
10.1016/j.ijmm.2013.02.013
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发表时间：
2013-05-01
影响因子：
4.1
通讯作者：
McClelland, Michael
中科院分区：
文献类型：
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作者：
Chandra, Mudit;Cheng, Pui;McClelland, Michael
大肠杆菌通常是共生菌，但也包括一些在质粒和/或染色体中携带额外基因的高致病性菌株。根据这些基因将病原菌分为肠致病性(EPEC)、肠出血性(EHEC)、产毒性(ETEC)、聚集性(EAEC)、侵袭性(EIEC)和弥漫性粘附型(DAEC)。在这里，以前开发的针对这些菌株的多重PCR策略被集成到一个单一步骤的多重策略中，该策略通常基于多个特征的PCR产物来区分所有这些大肠杆菌致病类型。该多重聚合酶链式反应适用于菌落聚合酶链式反应。添加了两个额外的标记：一个用于检测大多数肠杆菌科细菌，作为成功聚合酶链式反应的阳性对照，另一个用于区分沙门氏菌。菌落聚合酶链式反应对45株参比菌株进行了正确的分类，丝素聚合酶链式反应对71株(45+26)菌株进行了正确分类。然后用它来询问来自牛宿主的44个临床菌株，从而检测EAEC和DAEC决定簇。(C)2013年爱思唯尔股份有限公司。版权所有。
E. coli is generally a commensal but includes some highly pathogenic strains carrying additional genes in plasmids and/or the chromosome. Based on these genes the pathogenic strains are divided into pathotypes including enteropathogenic (EPEC), enterohemorrhagic (EHEC), enterotoxigenic (ETEC), enteroaggregative (EAEC), enteroinvasive (EIEC) and diffusely adherent (DAEC) E. coli. Here, previously developed multiplex PCR strategies for these strains were integrated into one single step multiplex that differentiates all these E. coli pathotypes, usually based on multiple characteristic PCR products. This multiplex PCR works reliably for colony PCR. Two additional markers were added: one to detect most Enterobacteriacea, which acts as a positive control for successful PCR, and one to distinguish Salmonella. The multiplex correctly classified a set of 45 reference strains by colony PCR and 71 (45 + 26) strains by in silk PCR. It was then used to interrogate 44 clinical strains from bovine hosts resulting in detection of EAEC and DAEC determinants. (C) 2013 Elsevier GmbH. All rights reserved.