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Development of a new microporous filter method for the concentration of viruses from water.

机译：开发了一种新的微孔过滤器方法，用于从水中浓缩病毒。

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摘要

Waterborne enteric viruses are transmitted via the fecal-oral route and have been isolated from various types of water ranging from sewage to tap water. Water matrices characterized by low levels of organic material (e.g. clean surface water and tap water) contain fewer numbers of viruses than sewage and wastewater effluents. A number of methods have been developed to concentrate, elute (recover), and re-concentrate viruses from water. The goal of this dissertation is two-fold. An extensive review of the literature is provided in Appendix A that focuses on method development in the three aforementioned areas. A review of this detail has not been conducted in over two decades, and as such will contribute to the fields of water quality and environmental virology. Second, a novel and inexpensive method for the concentration of viruses (MS2 coliphage, poliovirus 1, echovirus 1, Coxsackievirus B5, and adenovirus 2) is presented in Appendix B. The method uses a new electropositive filter (comprised of nanoalumina fibers) for the capture of viruses from 20-L volumes of dechlorinated tap water. Average filter retention efficiencies for each of the viruses was ≥ 99%. Viruses that are adsorbed to filters must then be recovered (eluted). A number of inorganic solutions were evaluated for this purpose, the most effective being a moderately alkaline (pH 9.3) glycine buffered-polyphosphate solution. Secondary reconcetration of the eluates was performed using an optimized ultrafiltration method (Centricon Plus-70, Millipore, Billerica, MA), and achieved final concentrates volumes of 3.3 +/- 0.3 mL. Total method efficiencies meeting the project recovery goal of ≥ 50% were obtained for each of the tested viruses except for MS2 coliphage at high input titers (45 +/- 15%) and adenovirus 2 (14 +/- 4%). Appendix C provides the Standard Operating Procedures, sample calculations, and detailed data for the experiments conducted. Appendix D details the steps taken towards optimizing the secondary concentration procedure in effort to meet the 50% recovery goal.
机译：水性肠道病毒通过粪-口途径传播，并已从污水到自来水等各种类型的水中分离出来。以有机物含量较低（例如，干净的地表水和自来水）为特征的水基质所含的病毒数量要少于污水和废水。已经开发了许多方法来从水中浓缩，洗脱（回收）和重新浓缩病毒。本文的目的是双重的。附录A中提供了对文献的详尽回顾，重点是上述三个领域中的方法开发。在过去的二十多年中尚未对此细节进行审查，因此将有助于水质和环境病毒学领域。其次，附录B中提供了一种新颖且廉价的病毒浓缩方法（MS2噬菌体，脊髓灰质炎病毒1，回声病毒1，柯萨奇病毒B5和腺病毒2）。该方法使用了一种新型的正电过滤器（由纳米氧化铝纤维组成）。从20升体积的脱氯自来水中捕获病毒。每种病毒的平均过滤器保留效率均≥99％。然后必须回收（洗脱）吸附在过滤器上的病毒。为此目的，评估了许多无机溶液，最有效的是中等碱性（pH 9.3）的甘氨酸缓冲多磷酸盐溶液。使用优化的超滤方法（Centricon Plus-70，Millipore，Billerica，MA）进行洗脱液的二次浓缩，最终浓缩液体积为3.3 +/- 0.3 mL。除了在高输入滴度（45 +/- 15％）和腺病毒2（14 +/- 4％）时的MS2噬菌体，每种测试病毒均获得了满足项目恢复目标≥50％的方法总效率。附录C提供了所进行实验的标准操作程序，样品计算和详细数据。附录D详细介绍了为达到50％回收率目标而优化二级浓缩程序所采取的步骤。

著录项

  • 作者

    Ikner, Luisa Antoinette.;

  • 作者单位

    The University of Arizona.;

  • 授予单位 The University of Arizona.;
  • 学科 Biology Microbiology.;Environmental Sciences.;Biology Virology.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 196 p.
  • 总页数 196
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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