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首页> 外文会议>International Conference on Environmental Science and Technology >CURRENT DEVELOPMENTS IN QUANTIFICATION OF PATHOGENS FROM ENVIRONMENTAL MEDIA: THE TRADEOFF OF ACCURACY AND EASE IN THE CASE OF SALMONELLA IN BIOSOLIDS
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CURRENT DEVELOPMENTS IN QUANTIFICATION OF PATHOGENS FROM ENVIRONMENTAL MEDIA: THE TRADEOFF OF ACCURACY AND EASE IN THE CASE OF SALMONELLA IN BIOSOLIDS

机译：来自环境媒体的病原体的目前的发展：在生物溶解中的沙门氏菌的准确性和易于易于易受的权衡

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The quantification of Salmonella in biosolids is typically performed by culture-based techniques, which are time-consuming and vulnerable to many confounding problems such as the underestimation of pathogen levels due to the presence of viable but non-culturable (VNBC) state cells. Molecular quantification techniques potentially offer much more rapid and reliable analysis. This study is aimed at adaptation and application of the most current molecular based techniques, Taqman assisted qPCR and loop-mediated isothermal amplification (LAMP) to this pressing public health issue. Equally important, by obtaining Salmonella counts by the best available culture-based technique in parallel, this project will offer reliable counts of Salmonella in Turkish biosolids in a manner that allows comparison to international numbers.A pressing issue regarding the application of molecular based methods for counting pathogens in biosolids is that non-viable DNA may be amplified by these techniques, which would in turn lead to over-estimation of cell counts. A recently developed and verified technique for distinguishing the genetic material in intact, viable cells from genetic material outside of cells or contained in mortally damaged cells has been developed and demonstrated. This technique, which is based on the binding of the chemical propidium monoazide (PMA) to extracellular DNA only, potentially offers the most rapid and reliable method for distinguishing viable and non-viable cells to date, thus potentially solving the final challenge that faces molecular Salmonella quantification. Recently digested biosolids will be collected from a handful of municipal sewage treatment works in Turkey. Salmonella in samples were analyzed in parallel by three methods: 1) USA EPA method 1682 for determination of most probable number by culture-based techniques, 2) quantitative PCR (qPCR) with application of propidium monoazide and Taqman probes and 3) loop-mediated DNA amplification with PMA treatment. Initial results of these studies will be presented.
机译：生物糖中的沙门氏菌的定量通常通过基于培养基的技术进行，这是耗时的，并且易于许多混淆问题，例如由于存在活性但非培养的（VNBC）状态细胞的存在而低估病原体水平。分子量化技术可能提供更快速可靠的分析。本研究旨在适应和应用最新的基于分子的技术，Taqman辅助QPCR和环介导的等温扩增（灯）到这一紧迫的公共卫生问题。同样重要的是，通过并行获得最佳培养的技术的奶士计数，该项目将以允许与国际数字进行比较的方式提供土耳其生物溶解体中的可靠计数。关于基于分子方法的施加的压制问题在生物溶胶中计数病原体是可以通过这些技术扩增不可活性DNA，这反过来又导致细胞计数的过度估计。最近开发和验证的技术，用于区分遗传物质的完整，可行的细胞从细胞外的遗传物质中或含有致命受损细胞中的遗传物质。这种技术基于化学促氧化物（PMA）与细胞外DNA的结合，可能提供了用于区分可行和不可行的细胞迄今为止的最快速和可靠的方法，因此可能解决了所分子的最终挑战沙门氏菌量化。最近消化的生物溶解将从土耳其的少数城市污水处理工程中收集。样品中的Salmonella以三种方法分析：1）美国EPA方法1682通过培养基技术，2）定量PCR（QPCR）测定最可能的数量，其中包含单氮杂甙和Taqman探针和3）环介导用PMA处理进行DNA扩增。将提出这些研究的初始结果。

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