Viral genome isolation from human faeces for succession assessment of the human gut virome

TY - CHAP

T1 - Viral genome isolation from human faeces for succession assessment of the human gut virome

AU - Mayo Munoz, David

PY - 2018

Y1 - 2018

N2 - Despite the important role of the microbiota in the human gastrointestinal tract (GIT) and its impact on life-long health, the successional process through which this microbial community develops during infancy is still poorly understood. Specially, little is known about how the amount and type of viruses present in the GIT, i.e., the virome, varies throughout this period and about the role this collection of viruses may play in the assembly of the GIT microbiota. The patterns of taxonomic change of the GIT viral community can be analyzed in a birth cohort of infants during the first year of life. The present chapter presents a detailed protocol for the isolation and extraction of viral nucleic acids from collected human faecal samples, whole genome amplification (WGA) using phi29 DNA polymerase and preparation for sequencing through high-throughput 454 pyrosequencing. The sequencing data can be posteriorly used for taxonomic classification in order to establish the composition of the virome present in each sample and to assess the process of viral dynamics through time.

AB - Despite the important role of the microbiota in the human gastrointestinal tract (GIT) and its impact on life-long health, the successional process through which this microbial community develops during infancy is still poorly understood. Specially, little is known about how the amount and type of viruses present in the GIT, i.e., the virome, varies throughout this period and about the role this collection of viruses may play in the assembly of the GIT microbiota. The patterns of taxonomic change of the GIT viral community can be analyzed in a birth cohort of infants during the first year of life. The present chapter presents a detailed protocol for the isolation and extraction of viral nucleic acids from collected human faecal samples, whole genome amplification (WGA) using phi29 DNA polymerase and preparation for sequencing through high-throughput 454 pyrosequencing. The sequencing data can be posteriorly used for taxonomic classification in order to establish the composition of the virome present in each sample and to assess the process of viral dynamics through time.

KW - GIT microbiota

KW - High-throughput sequencing

KW - Succession

KW - Viral genome extraction

KW - Virome

KW - Virus isolation

KW - Whole genome amplification

U2 - 10.1007/978-1-4939-8682-8_8

DO - 10.1007/978-1-4939-8682-8_8

M3 - Book chapter

C2 - 30128992

AN - SCOPUS:85052217161

SN - 978-1-4939-8681-1

T3 - Methods in Molecular Biology

SP - 97

EP - 108

BT - The Human Virome

A2 - Moya, Andrés

A2 - Brocal, Vicente Pérez

PB - Humana Press

ER -