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通过下一代测序技术对污水和粪便样本中的诺如病毒和其他人类肠道病毒进行特征分析。

Characterization of Norovirus and Other Human Enteric Viruses in Sewage and Stool Samples Through Next-Generation Sequencing.

机构信息

Ifremer, Laboratoire de Microbiologie, LSEM-SG2M, BP 21105, 44311, Nantes Cedex 3, France.

Department of Viroscience, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.

出版信息

Food Environ Virol. 2019 Dec;11(4):400-409. doi: 10.1007/s12560-019-09402-3. Epub 2019 Aug 24.

Abstract

This study aimed to optimize a method to identify human enteric viruses in sewage and stool samples using random primed next-generation sequencing. We tested three methods, two employed virus enrichment based on the binding properties of the viral capsid using pig-mucin capture or by selecting viral RNA prior to library preparation through a capture using the SureSelect target enrichment. The third method was based on a non-specific biophysical precipitation with polyethylene glycol. Full genomes of a number of common human enteric viruses including norovirus, rotavirus, husavirus, enterovirus and astrovirus were obtained. In stool samples full norovirus genome were detected as well as partial enterovirus genome. A variety of norovirus sequences was detected in sewage samples, with genogroup II being more prevalent. Interestingly, the pig-mucin capture enhanced not only the recovery of norovirus and rotavirus but also recovery of astrovirus, sapovirus and husavirus. Documenting sewage virome using these methods provides information for molecular epidemiology and may be useful in developing strategies to prevent further spread of viruses.

摘要

本研究旨在优化一种使用随机引物的下一代测序方法,以鉴定污水和粪便样本中的人类肠道病毒。我们测试了三种方法,两种方法基于病毒衣壳的结合特性使用猪粘蛋白捕获或通过在文库制备前通过使用 SureSelect 目标富集进行病毒 RNA 选择进行病毒富集。第三种方法基于非特异性的聚乙二醇物理沉淀。获得了许多常见的人类肠道病毒,包括诺如病毒、轮状病毒、肝炎病毒、肠病毒和星状病毒的全长基因组。在粪便样本中检测到了完整的诺如病毒基因组和部分肠病毒基因组。在污水样本中检测到了多种诺如病毒序列,其中 II 型诺如病毒更为普遍。有趣的是,猪粘蛋白捕获不仅增强了诺如病毒和轮状病毒的回收率,还增强了星状病毒、肠型杯状病毒和肝炎病毒的回收率。使用这些方法记录污水病毒组可提供分子流行病学信息,并可能有助于制定预防病毒进一步传播的策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23f3/6848244/3a47727e2945/12560_2019_9402_Fig1_HTML.jpg

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