Mayo-Muñoz David
Danish Archaea Centre, Department of Biology, University of Copenhagen, Copenhagen, Capital Region, Denmark.
Methods Mol Biol. 2018;1838:97-108. doi: 10.1007/978-1-4939-8682-8_8.
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.
尽管微生物群在人类胃肠道（GIT）中起着重要作用，并对终身健康产生影响，但人们对这一微生物群落在婴儿期的发育演替过程仍知之甚少。特别是，关于胃肠道中存在的病毒的数量和类型，即病毒组，在这一时期如何变化，以及这群病毒在胃肠道微生物群的组装中可能发挥的作用，人们了解得很少。可以在一组出生队列的婴儿生命的第一年中分析胃肠道病毒群落的分类学变化模式。本章介绍了从收集的人类粪便样本中分离和提取病毒核酸、使用phi29 DNA聚合酶进行全基因组扩增（WGA）以及通过高通量454焦磷酸测序进行测序准备的详细方案。测序数据随后可用于分类学分类，以确定每个样本中存在的病毒组的组成，并评估病毒随时间的动态变化过程。
