Extraction and purification of viruses from fecal samples for metagenome and morphology analyses

Josué L. Castro-Mejía*, Ling Deng, Finn K. Vogensen, Alejandro Reyes, Dennis S. Nielsen

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

6 Citations (Scopus)

Abstract

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.

Original languageEnglish
Title of host publicationThe Human Virome : Methods and Protocols
EditorsAndrés Moya, Vicente Pérez Brocal
Number of pages9
PublisherHumana Press
Publication date2018
Pages49-57
ISBN (Print) 978-1-4939-8681-1
ISBN (Electronic) 978-1-4939-8682-8
DOIs
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
Volume1838
ISSN1064-3745

Keywords

  • Extraction procedures
  • Gut-virome
  • Metagenome of extracellular viral-like particles (eVLPs)
  • Morphology
  • Polyethylene-glycol (PEG) precipitation

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