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

David Mayo Munoz

Abstract

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.

Original languageEnglish
Title of host publicationThe Human Virome : Methods and Protocols
EditorsAndrés Moya, Vicente Pérez Brocal
Number of pages12
PublisherHumana Press
Publication date2018
Pages97-108
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

  • GIT microbiota
  • High-throughput sequencing
  • Succession
  • Viral genome extraction
  • Virome
  • Virus isolation
  • Whole genome amplification

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