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首页> 外文期刊>Journal of Clinical Microbiology >Specific Detection of Enteroaggregative Hemorrhagic Escherichia coli O104:H4 Strains by Use of the CRISPR Locus as a Target for a Diagnostic Real-Time PCR
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Specific Detection of Enteroaggregative Hemorrhagic Escherichia coli O104:H4 Strains by Use of the CRISPR Locus as a Target for a Diagnostic Real-Time PCR

机译：通过使用CRISPR基因位点作为诊断实时PCR的靶标特异性检测肠聚合性出血性大肠杆菌O104：H4菌株

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摘要

In 2011, a large outbreak of an unusual bacterial strain occurred in Europe. This strain was characterized as a hybrid of an enteroaggregative Escherichia coli (EAEC) and a Shiga toxin-producing E. coli (STEC) strain of the serotype O104:H4. Here, we present a single PCR targeting the clustered regularly interspaced short palindromic repeats locus of E. coli O104:H4 (CRISPRO104:H4) for specific detection of EAEC STEC O104:H4 strains from different geographical locations and time periods. The specificity of the CRISPRO104:H4 PCR was investigated using 1,321 E. coli strains, including reference strains for E. coli O serogroups O1 to O186 and flagellar (H) types H1 to H56. The assay was compared for specificity using PCR assays targeting different O104 antigen-encoding genes (wbwCO104, wzxO104, and wzyO104). The PCR assays reacted with all types of E. coli O104 strains (O104:H2, O104:H4, O104:H7, and O104:H21) and with E. coli O8 and O9 strains carrying the K9 capsular antigen and were therefore not specific for detection of the EAEC STEC O104:H4 type. A single PCR developed for the CRISPRO104:H4 target was sufficient for specific identification and detection of the 48 tested EAEC STEC O104:H4 strains. The 35 E. coli O104 strains expressing H types other than H4 as well as 8 E. coli strains carrying a K9 capsular antigen tested all negative for the CRISPRO104:H4 locus. Only 12 (0.94%) of the 1,273 non-O104:H4 E. coli strains (serotypes Ont:H2, O43:H2, O141:H2, and O174:H2) reacted positive in the CRISPRO104:H4 PCR (99.06% specificity).
机译：2011年，欧洲发生了一次不寻常细菌菌株的大规模爆发。该菌株的特征是肠道聚集性大肠杆菌（EAEC）和血清型O104：H4的产志贺毒素的大肠杆菌（STEC）菌株的杂种。在这里，我们提出了一个针对以 c 聚簇的 r （通常是 i n个间隔的 s hort p < / em> alindromic r 重复检测大肠杆菌O104：H4（CRISPR O104：H4 ）的基因座，可从不同地理位置和时间特异性检测EAEC STEC O104：H4菌株期。使用1,321株E. coli菌株研究了CRISPR O104：H4 PCR的特异性，包括E. coli O血清群O1至O186和鞭毛（H）类型H1至H56的参考菌株。使用针对不同O104抗原编码基因（ wbwC O104 ， wzx O104 ，以及 wzy O104 ）。 PCR分析与所有类型的大肠杆菌O104菌株（O104：H2，O104：H4，O104：H7和O104：H21）以及与带有K9荚膜抗原的大肠杆菌O8和O9菌株反应，因此没有特异性用于检测EAEC STEC O104：H4类型。为CRISPR O104：H4 目标开发的单个PCR足以特异性鉴定和检测48种测试的EAEC STEC O104：H4菌株。 35个表达H4而非H4的大肠杆菌O104菌株以及8个带有K9荚膜抗原的大肠杆菌对CRISPR O104：H4 基因座均呈阴性。在1,273个非O104：H4大肠杆菌菌株（血清型Ont：H2，O43：H2，O141：H2和O174：H2）中只有12个（0.94％）在CRISPR O104：H4中反应呈阳性 PCR（99.06％特异性）。

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