Abstract：
:A major limitation for better understanding the role of the human gut virome in health and disease is the lack of validated methods that allow high throughput virome analysis. To overcome this, we evaluated the quantitative effect of homogenisation, centrifugation, filtration, chloroform treatment and random amplification on a mock-virome (containing nine highly diverse viruses) and a bacterial mock-community (containing four faecal bacterial species) using quantitative PCR and next-generation sequencing. This resulted in an optimised protocol that was able to recover all viruses present in the mock-virome and strongly alters the ratio of viral versus bacterial and 16S rRNA genetic material in favour of viruses (from 43.2% to 96.7% viral reads and from 47.6% to 0.19% bacterial reads). Furthermore, our study indicated that most of the currently used virome protocols, using small filter pores and/or stringent centrifugation conditions may have largely overlooked large viruses present in viromes. We propose NetoVIR (Novel enrichment technique of VIRomes), which allows for a fast, reproducible and high throughput sample preparation for viral metagenomics studies, introducing minimal bias. This procedure is optimised mainly for faecal samples, but with appropriate concentration steps can also be used for other sample types with lower initial viral loads.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Conceição-Neto N,Zeller M,Lefrère H,De Bruyn P,Beller L,Deboutte W,Yinda CK,Lavigne R,Maes P,Van Ranst M,Heylen E,Matthijnssens Jdoi
10.1038/srep16532subject
Has Abstractpub_date
2015-11-12 00:00:00pages
16532issn
2045-2322pii
srep16532journal_volume
5pub_type
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