A multiplex PCR method to detect 14 Escherichia coli serogroups associated with urinary tract infections
A multiplex PCR method to detect 14 Escherichia coli serogroups associated with urinary tract infections
复制标题
多重PCR方法检测与尿路感染相关的14种大肠杆菌血清群
DOI：
10.1016/j.mimet.2010.04.008
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发表时间：
2010-07-01
影响因子：
2.2
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Li, Dan;Liu, Bin;Wang, Lei
尿路感染(UTIs)是最常见的细菌感染之一，主要由尿路致病性大肠埃希菌(UPEC)引起。大肠杆菌属于14个血清群，包括O1、O2、O4、O6、O7、O8、O15、O16、O18、O21、O22、O25。O75和O83是在各种临床尿液标本中最常检测到的UPEC菌株。在本研究中，我们对O1、O2、O18和O75群的O抗原基因簇进行了研究。建立了一种基于O抗原特异性基因的多重聚合酶链式反应方法，可同时检测所有14个大肠杆菌血清群。对186株大肠埃希菌和志贺氏菌群参考菌株、47株大肠埃希菌临床分离株和10株其他菌种的检测表明，多重聚合酶链式反应方法具有高度的特异性和重复性。分析了多重聚合酶链式反应方法检测O抗原特异性基因的敏感性，并用该方法对5例医院尿液标本进行了检测，其中1例用常规的血清分型方法进行了验证。本文开发的多重聚合酶链式反应方法可用于检测临床和/或环境样本中的相关大肠杆菌菌株，尤其适用于尿路感染患者的尿样流行病学分析。(C)2010爱思唯尔公司版权所有。
Urinary tract infections (UTIs) are one of the most common bacterial infections and are predominantly caused by uropathogenic Escherichia coli (UPEC). E. coli strains belonging to 14 serogroups, including O1, O2, O4, O6, O7, O8, O15, O16, O18, O21, O22, O25. O75 and O83, are the most frequently detected UPEC strains in a diverse range of clinical urine specimens. In the current study, the O-antigen gene clusters of E. coli serogroups O1, O2, O18 and O75 were characterized. A multiplex PCR method based on O-antigen-specific genes was developed for the simultaneous detection of all 14 E. coli serogroups. The multiplex PCR method was shown to be highly specific and reproducible when tested against 186 E. coli and Shigella O-serogroup reference strains, 47 E. coli clinical isolates and 10 strains of other bacterial species. The sensitivity of the multiplex PCR method was analyzed and shown to detect O-antigen-specific genes in samples containing 25 ng of genomic DNA or in mock urine specimens containing 40 colony-forming units (CFUs) per ml. Five urine specimens from hospital were examined using this multiplex PCR method, and the result for one sample was verified by the conventional serotyping methods. The multiplex PCR method developed herein can be used for the detection of relevant E. coli strains from clinical and/or environmental samples, and it is particularly useful for epidemiologic analysis of urine specimens from patients with UT's. (C) 2010 Elsevier B.V. All rights reserved.