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>Differentiation of stx1A gene for detection of Escherichia coli serotype O157: H7 and Shigella dysenteriae type 1 in food samples using high resolution melting curve analysis
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Differentiation of stx1A gene for detection of Escherichia coli serotype O157: H7 and Shigella dysenteriae type 1 in food samples using high resolution melting curve analysis
Escherichia coli serotype O157: H7 and Shigella dysenteriae type 1 as the Shiga toxin‐producing bacteria cause some acute gastrointestinal and extraintestinal diseases such as hemorrhagic uremic syndrome and bloody diarrhea in human. Stx genes are the key virulence factors in these pathogens. The aim of this study was to develop HRMA assay to differentiate stx1A gene for detection of E. coli serotype O157: H7 and Sh. dysenteriae type 1 and determine the prevalence of these pathogens in food samples using this method. PCR‐HRMA assay and gold standard methods have been carried out for identification of pathogens among 135 different food samples. We found HRMA method a sensitive and specific assay (100 and 100%, respectively) for differentiation of stx1A gene, consequently, detection of these pathogens in food samples. Also, the highest prevalence of E. coli serotype O157: H7 and Sh. dysenteriae type 1 harboring stx1A gene was observed in raw milk and vegetable salad samples, respectively. HRMA as a rapid, inexpensive, sensitive and specific method is suggested to be used for differentiation of stx1A gene to detect E. coli serotype O157: H7 and Sh. dysenteriae type 1 as the key pathogens for safety evaluation of food samples.
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机译：大肠杆菌血清型O157：H7和Shigella Dysenteriae Type 1作为滋阴产生的细菌，导致一些急性胃肠和肾外疾病，如出血性尿毒症综合征和人类的血腥腹泻。 STX基因是这些病原体中的关键毒力因子。本研究的目的是发展HRMA测定以区分STX1A基因用于检测大肠杆菌血清型O157：H7和SH。痢疾1型，并使用该方法确定食品样品中这些病原体的患病率。已经进行了PCR-HRMA测定和黄金标准方法，用于鉴定135种不同的食物样品中病原体。我们发现HRMA方法对于STX1A基因的分化，因此，在食物样品中检测这些病原体的敏感和特异性测定（分别为100和100％）。此外，大肠杆菌血清型O157：H7和Sh的最高普遍率。在原料牛奶和蔬菜沙拉样品中观察到吞咽STX1A基因的缺乏亢进型1型。建议使用HRMA作为快速，廉价，灵敏和特异性的方法，用于分化STX1A基因以检测大肠杆菌血清型O157：H7和SH。 Dysenteriae Type 1作为食物样品安全评估的关键病原体。
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