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首页> 外文期刊>Food analytical methods >The DNA sequence of the Escherichia coli O22 O-antigen gene cluster and detection of pathogenic strains belonging to E. coli serogroups O22 and O91 by multiplex PCR assays targeting virulence genes and genes in the respective O-antigen gene clusters.
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The DNA sequence of the Escherichia coli O22 O-antigen gene cluster and detection of pathogenic strains belonging to E. coli serogroups O22 and O91 by multiplex PCR assays targeting virulence genes and genes in the respective O-antigen gene clusters.

机译：大肠杆菌O22 O-抗原基因簇的DNA序列和属于 E的致病菌的检测。通过针对毒力基因和各个O抗原基因簇中的基因的多重PCR分析，确定O22和O91血清型。

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Multiplex polymerase chain reaction (PCR) assays were developed for detection of pathogenic strains belonging to Escherichia coli serogroups O22 and O91. The O-antigen gene cluster of E. coli O22 was sequenced to identify genes that could be employed as targets for serogroup-specific PCR assays. The wzx and wzy genes in the O-antigen gene clusters of E. coli O22 and E. coli O91 were selected as target genes. The assays were serogroup-specific when tested against 72 E. coli O22 strains and 57 E. coli O91 strains isolated from food, humans, and animals, representative strains belonging to 168 E. coli O serogroups and non-E. coli bacteria. Furthermore, 72 E. coli O22 strains and 57 E. coli O91 strains isolated from food, water, animals, and humans were tested by the PCR for the presence of six and 19 virulence genes, respectively, associated with pathogenic E. coli strains. Based on the PCR screening results, multiplex PCR assays targeting the O22 wzy gene and the cnf-1 and sfa genes in E. coli O22 and the O91 wzy gene, conserved sequences of stx1 and stx2 genes, and the astA and cdt-III genes in E. coli O91 were developed to detect and identify pathogenic strains belonging to serogroups O22 and O91. Furthermore, E. coli O22 and O91 were detected by multiplex PCR assays targeting the wzx or wzy genes and conserved sequences of the stx1 and stx2 genes in ground beef samples inoculated with approximately two colony-forming units (CFU)/25 g after 18-h enrichment. The results demonstrate that the E. coli O22 and O91 wzx and wzy gene sequences were specific for the respective serogroups and can be used as diagnostic markers for rapid identification of these serogroups as an alternative to serotyping. The multiplex PCR assays targeting the O22 and O91 wzx and wzy genes and virulence genes can be used to identify and to detect pathogenic strains of these serogroups in food and fecal samples.
机译：开发了多重聚合酶链反应（PCR）检测试剂盒，用于检测大肠杆菌（Escherichia coli）血清型O22和O91的致病菌株。 E的O-抗原基因簇。对大肠杆菌O22进行测序，以鉴定可用作血清群特异性PCR分析靶标的基因。 E的O抗原基因簇中的 wzx 和 wzy 基因。大肠杆菌 O22和 E。选择大肠杆菌O91作为靶基因。当针对72 E进行测试时，该测定是血清群特异性的。大肠杆菌O22菌株和57种大肠杆菌。从食物，人和动物中分离出的大肠杆菌O91菌株，属于168 E的代表性菌株。大肠杆菌O血清群和非 E。大肠菌。此外，为72E。大肠杆菌O22菌株和57种大肠杆菌。通过PCR检测了从食物，水，动物和人分离出的大肠杆菌O91菌株中分别存在与致病性大肠杆菌相关的6个和19个毒力基因。大肠杆菌菌株。根据PCR筛选结果，以O22 wzy 基因以及 E中的 cnf -1和 sfa 基因为目标的多重PCR分析。大肠杆菌 O22和O91 wzy 基因， stx 1 和 stx 的保守序列2 基因，以及 E中的 astA 和 cdt -III基因。开发了O91大肠埃希氏菌以检测和鉴定O22和O91血清群的致病菌。此外， E。通过针对 wzx 或 wzy 基因和 stx 保守序列的多重PCR分析检测到O22和O91富集18小时后，用大约两个菌落形成单位（CFU）/ 25 g接种的碎牛肉样品中的1 和 stx 2 基因。结果表明 E。大肠杆菌O22和O91 wzx 和 wzy 基因序列对各自的血清群具有特异性，可以用作快速鉴定这些血清群的诊断标志物，以替代分型。针对O22和O91 wzx 和 wzy 基因和毒力基因的多重PCR分析可用于识别和检测食品和粪便样品中这些血清群的致病菌株。

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