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Helicobacter Pylori Detection in Shellfish: A Real-Time Quantitative Polymerase Chain Reaction Approach

机译:贝类幽门螺杆菌检测:实时定量聚合酶链反应方法

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摘要

Helicobacter pylori emerges as one of the most concerning human pathogens associated to water and, thereby, it could be present in raw and slightly treated marine food products. The present research work aimed to detect the presence of H. pylori in Spanish commercial samples of shellfish (mussels, clams, and cockles) by means of a quantitative real-time polymerase chain reaction (qPCR) approach based on the vacuolating cytotoxin A ( vacA ) gene specificity. Putative H. pylori amplicons were confirmed by sequencing. qPCR was positive for 12 out of the 100 samples, being 67% (8/12) from mussels, 25% (3/12) from clams, and only 8% (1/12) from cockles. After sequencing, three of the amplicons showed 97–99% homology with the H. pylori vacA gene. Quantitative results indicate that the levels of contamination remained below 10~(2) log_(10) colony forming units per mL (CFU/mL). The present research shows for the first time the effectiveness of the optimized qPCR in the identification of potentially H. pylori contaminated shellfish products. Our results confirm the presence of H. pylori in shellfish from the Spanish western seacoast and verify the possible relationship between the presence of H. pylori in seawater and the role of contaminated seafoods as vehicles of H. pylori entrance into the food chain.
机译:幽门螺杆菌出现是与水相关的人类病原体中最多的一种,从而可以存在于原料和略微处理的海洋食品中。目前的研究工作旨在通过定量的实时聚合酶链反应(QPCR)方法检测西班牙商业样品H. Pylori的存在。基于真空的细胞毒素A(Vaca )基因特异性。通过测序证实了幽门螺杆菌扩增子。 QPCR为100个样品中的12种阳性,来自贻贝的67%(8/12),来自蛤蜊的25%(3/12),距离蛤蜊仅为8%(1/12)。测序后,三个扩增子显示出97-99%的同源性与H. Pylori Vaca基因。定量结果表明,污染水平仍然低于10〜(2)LOG_(10)菌落形成单位(CFU / mL)。本研究表明,首次优化QPCR在识别潜在的H.幽门污染的贝类产品中的有效性。我们的结果证实了来自西班牙西部海岸的贝类中H. Pylori的存在,并核实海水中H. Pylori的存在与污染的海鲜作为H. Pylori入口进入食物链中的杂草的作用。

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