Castro-Mejía Josué L, Deng Ling, Vogensen Finn K, Reyes Alejandro, Nielsen Dennis S
Department of Food Science, University of Copenhagen, Copenhagen, Denmark.
Department of Biological Sciences, Universidad de los Andes, Bogotá, Colombia.
Methods Mol Biol. 2018;1838:49-57. doi: 10.1007/978-1-4939-8682-8_5.
The human enteric virome consists of endogenous retro elements and viruses that infect the host and members of the gut microbiome (GM). Mounting evidence suggests that the gut virome plays a central role in maintaining homeostasis and via the GM influences immunology of the host. To thoroughly characterize the gut virome, it is often very useful to first separate and concentrate extracellular viral-like particles (eVLPs) enabling an integrative characterization of them. Here, we describe a detailed protocol for extraction and concentration of the viral fraction from fecal samples based on a polyethylene glycol precipitation (PEG) approach. These procedures maximize the yields of eVLPs (and their DNA) with high purity well suited for down-stream analysis such as quantification and morphological assessment, determination of phage-host pairs as well as virome sequencing.
人类肠道病毒组由内源性逆转录元件以及感染宿主和肠道微生物群（GM）成员的病毒组成。越来越多的证据表明，肠道病毒组在维持体内平衡中起核心作用，并通过肠道微生物群影响宿主的免疫学。为了全面表征肠道病毒组，首先分离和浓缩细胞外病毒样颗粒（eVLP）通常非常有用，这有助于对它们进行综合表征。在这里，我们描述了一种基于聚乙二醇沉淀（PEG）方法从粪便样本中提取和浓缩病毒组分的详细方案。这些程序最大限度地提高了eVLP（及其DNA）的产量，其高纯度非常适合下游分析，如定量和形态评估、噬菌体-宿主对的确定以及病毒组测序。
