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首页> 外文期刊>Food microbiology >Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization
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Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization

机译：致病性耶尔森氏菌的差异检测。通过荧光原位杂交

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摘要

Yersinia enterocolitica, Y. pseudotuberculosis and Y. pestis are pathogens of major medical importance, which are responsible for a considerable number of infections every year. The detection of these species still relies on cultural methods, which are slow, labour intensive and often hampered by the presence of high amounts of accompanying flora. In this study, fluorescence in situ hybridization (FISH) was used to develop a fast, sensitive and reliable alternative to detect viable bacteria in food. For this purpose, highly specific probes targeting the 16S and 23S ribosomal RNA were employed to differentially detect each of the three species. In order to enable the differentiation of single nucleotide polymorphisms (SNPs), suitable competitor oligonucleotides and locked nucleic acids (LNAs) were used. Starved cells still showed a strong signal and a direct viable count (DVC) approach combined with FISH optimized live/ dead discrimination. Sensitivity of the FISH test was high and even a single cell per gram of spiked minced pork meat could be detected within a day, demonstrating the applicability to identify foodborne hazards at an early stage. In conclusion, the established FISH tests proved to be promising tools to compensate existing drawbacks of the conventional cultural detection of these important zoonotic agents.
机译：小肠结肠炎耶尔森氏菌，假结核耶尔森氏菌和鼠疫耶尔森氏菌是具有重要医学意义的病原体，每年引起大量感染。对这些物种的检测仍依赖于培养方法，该方法缓慢，劳动强度大，并且经常因存在大量伴随的菌群而受阻。在这项研究中，荧光原位杂交（FISH）用于开发一种快速，灵敏和可靠的替代方法来检测食品中的活菌。为此，采用靶向16S和23S核糖体RNA的高特异性探针来差异检测这三个物种。为了能够区分单核苷酸多态性（SNP），使用了合适的竞争寡核苷酸和锁核酸（LNA）。饥饿的细胞仍然显示强信号和直接活计数（DVC）方法，并结合FISH优化的活/死鉴别。 FISH测试的灵敏度很高，甚至在一天之内甚至可以检测到每克加碎猪肉中的单个细胞，这证明了其在早期发现食源性危害的适用性。总之，已建立的FISH测试被证明是有前途的工具，可以弥补这些重要的人畜共患病菌的常规文化检测所存在的缺陷。

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  • 来源
    《Food microbiology》 |2017年第4期|39-45|共7页
  • 作者

    Alexander Rohde; Jens Andre Hammerl; Bernd Appel; Ralf Dieckmann; Sascha Al Dahouk;

  • 作者单位

    German Federal Institute for Risk Assessment, Department of Biological Safety, Diedersdorfer Weg 1, D-12277 Berlin, Germany,Free University Berlin, Department of Biology, Chemistry and Pharmacy, Takustr. 3, 14195 Berlin, Germany;

    German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany;

    German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany;

    German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany;

    German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Yersinia; FISH; Food safety; Direct viable count;

    机译：耶尔森氏菌;鱼;食品安全;直接可行计数;

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