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Multiplex real-time PCR assay for detection of Escherichia coli O157:H7 and screening for non-O157 Shiga toxin-producing E. coli

机译:多重实时荧光定量PCR检测大肠杆菌O157:H7并筛选产非O157志贺毒素的大肠杆菌

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摘要

BackgroundShiga toxin-producing Escherichia coli (STEC), including E. coli O157:H7, are responsible for numerous foodborne outbreaks annually worldwide. E. coli O157:H7, as well as pathogenic non-O157:H7 STECs, can cause life-threating complications, such as bloody diarrhea (hemolytic colitis) and hemolytic-uremic syndrome (HUS). Previously, we developed a real-time PCR assay to detect E. coli O157:H7 in foods by targeting a unique putative fimbriae protein Z3276. To extend the detection spectrum of the assay, we report a multiplex real-time PCR assay to specifically detect E. coli O157:H7 and screen for non-O157 STEC by targeting Z3276 and Shiga toxin genes (stx1 and stx2). Also, an internal amplification control (IAC) was incorporated into the assay to monitor the amplification efficiency.
机译:背景技术产生志贺毒素的大肠杆菌(STEC),包括大肠杆菌O157:H7,是导致全球每年大量食源性疾病爆发的原因。大肠杆菌O157:H7以及病原性非O157:H7 STEC可能会导致危及生命的并发症,例如血性腹泻(溶血性结肠炎)和溶血性尿毒症综合征(HUS)。以前,我们开发了一种实时PCR分析法,通过靶向独特的假定菌毛蛋白Z3276来检测食品中的O157:H7大肠杆菌。为了扩展检测的检测范围,我们报告了一种多重实时PCR检测,以特异性检测大肠杆菌O157:H7,并通过靶向Z3276和志贺毒素基因(stx1和stx2)筛选非O157 STEC。另外,将内部扩增对照(IAC)掺入测定中以监测扩增效率。

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