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 af dette arbejde
3 Citationer (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.

OriginalsprogEngelsk
TitelThe Human Virome : Methods and Protocols
RedaktørerAndrés Moya, Vicente Pérez Brocal
Antal sider9
ForlagHumana Press
Publikationsdato2018
Sider49-57
ISBN (Trykt) 978-1-4939-8681-1
ISBN (Elektronisk) 978-1-4939-8682-8
DOI
StatusUdgivet - 2018
NavnMethods in Molecular Biology
Vol/bind1838
ISSN1064-3745

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