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首页> 外文期刊>Environmental Science & Technology >Combination of Crossflow Ultrafiltration, Monolithic Affinity Filtration, and Quantitative Reverse Transcriptase PCR for Rapid Concentration and Quantification of Model Viruses in Water
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Combination of Crossflow Ultrafiltration, Monolithic Affinity Filtration, and Quantitative Reverse Transcriptase PCR for Rapid Concentration and Quantification of Model Viruses in Water

机译：错流超滤，整体亲和过滤和定量逆转录酶PCR的组合，用于快速浓缩和定量水中的模型病毒

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摘要

We present a rapid and effective adsorption- elution method based on monolithic affinity filtration (MAF) for the concentration and purification of waterborne viruses. The MAF column consists of a hydrolyzed macroporous epoxy-based polymer. High recoveries were achieved by columns for the bacterial virus (bacteriophage) MS2 110 (±19)%, as model organism, as well as for human adenoviruses 42.4 (±3.4)% and murine noroviruses 42.6 (±1.9)%. This new concentration and purification method was combined with crossflow ultrafiltration (CUF). Because of the adsorption of the examined viruses to the macroporous surface of the MAF column at pH 3, concentrated matrix components by CUF can be removed. Bacteriophages MS2 were spiked in tap water and concentrated with the new CUF-MAF concentration method by a volumetric factor of 10~4 within 33 min. Furthermore, the detection limit for quantification of bacteriophage MS2 by quantitative reverse transcriptase PCR (qRT-PCR) could be improved from 79.47 to 0.0056 GU mL~(-1) by a factor of 1.4 × 10~4. In a first study, we have shown that this method could also be applied for river water containing naturally MS2 and MS2-like phages.
机译：我们提出了一种基于整体亲和过滤（MAF）的快速有效吸附吸附方法，用于浓缩和纯化水性病毒。 MAF柱由水解的大孔环氧基聚合物组成。通过色谱柱可对作为模型生物的细菌病毒（噬菌体）MS2 110（±19）％，人腺病毒42.4（±3.4）％和鼠诺如病毒42.6（±1.9）％的色谱柱实现高回收率。这种新的浓缩和纯化方法与错流超滤（CUF）相结合。由于所检查的病毒在pH 3时吸附到MAF色谱柱的大孔表面，因此可以通过CUF去除浓缩的基质成分。将噬菌体MS2掺入自来水中，并用新的CUF-MAF浓缩方法在33分钟内浓缩10〜4的体积因子。此外，通过定量逆转录酶PCR（qRT-PCR）进行噬菌体MS2定量检测的限度可以从79.47提高到0.0056 GU mL〜（-1）1.4×10〜4。在第一个研究中，我们表明该方法也可以用于含有天然MS2和MS2样噬菌体的河水中。

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  • 来源
    《Environmental Science & Technology》 |2012年第18期|p.10073-10080|共8页
  • 作者

    Lu Pei; Martin Rieger; Sandra Lengger; Sonja Ott; Claudia Zawadsky; Nils Marten Hartmann; Hans-Christoph Selinka; Andreas Tiehm; Reinhard Niessner; Michael Seidel;

  • 作者单位

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

    DVGW-Technologiezentrum Wasser, Department Environmental Biotechnology, Karlsruher Strafie 84, 76139 Karlsruhe, Germany;

    German Federal Environment Agency (UmweUbundesamt, UBA), FG II 1.4 Microbiological Risks, Corrensplatz 1, 14195 Berlin,Germany;

    German Federal Environment Agency (UmweUbundesamt, UBA), FG II 1.4 Microbiological Risks, Corrensplatz 1, 14195 Berlin,Germany;

    DVGW-Technologiezentrum Wasser, Department Environmental Biotechnology, Karlsruher Strafie 84, 76139 Karlsruhe, Germany;

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

    Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universitat Munchen, Marchioninistr. 17, 81377 Munich, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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