A Method for Isolation of the Virome from Plasma Samples

  • Pilar Domingo-CalapEmail author
  • Juan Vicente Bou
Part of the Methods in Molecular Biology book series (MIMB, volume 1838)


Virome studies are of special interest nowadays. Understanding viral communities in different body compartments will help guide future personalized treatments and to discern between homeostasis and disease. High-throughput sequencing technologies allow us to detect all the nucleic acids present in a sample, including viral ones, by random sequencing. One of the major challenges in virome studies is the correct isolation of the viral nucleic acids from a specific sample. This can be done during the extraction steps (e.g., enrichment of viral capsids), or during the bioinformatic analysis (e.g., removing all human and bacterial sequences). Furthermore, it is an important remark that the treatment of the sample will strongly influence the results. Samples will be treated differently if the ultimate goal is the study of all replicating and encapsidated viruses, including both RNA and DNA ones, if we are only focused on DNA ones, or if we want to analyze all the possible viral nucleic acids in the specific sample, even if the genome is degraded. Here, we present a technique that allows for isolation of viral nucleic acids from plasma samples.

Key words

Virus Plasma Isolation Nucleic acids 


  1. 1.
    Suttle CA (2007) Marine viruses--major players in the global ecosystem. Nat Rev Microbiol 5:801–812CrossRefPubMedGoogle Scholar
  2. 2.
    De Vlaminck I, Khush KK, Strehl C et al (2013) Temporal response of the human virome to immunosuppression and antiviral therapy. Cell 155:1178–1187CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kohl C, Brinkmann A, Dabrowski PW et al (2015) Protocol for metagenomic virus detection in clinical specimens. Emerg Infect Dis 21:48–57CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Pina S, Puig M, Lucena F et al (1998) Viral pollution in the environment and in shellfish: human adenovirus detection by PCR as an index of human viruses. Appl Environ Microbiol 64:3376–3382PubMedPubMedCentralGoogle Scholar
  5. 5.
    Conceição-Neto N, Zeller M, Lefrère H et al (2015) Modular aproach to customise sample preparation procedures for viral metagenomics: a reproducible protocol for virome analysis. Sci Rep 5:16532CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Zou N, Ditty S, Li B et al (2003) Random priming PCR strategy to amplify and clone trace amounts of DNA. BioTechniques 35:503–508Google Scholar
  7. 7.
    Victoria JG, Kapoor A, Dupuis K et al (2008) Rapid identification of known and new RNA viruses from animal tissues. PLoS Pathog 4:e1000163CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Hang J, Forshey BM, Kochel TJ et al (2012) Random amplification and pyrosequencing for identification of novel viral genome sequences. J Biomol Tech 23:4–10CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Evans GE, Murdoch DR, Anderson TP et al (2003) Contamination of Qiagen DNA extraction kits with legionella DNA. J Clin Virol 41:3452–3453Google Scholar
  10. 10.
    Erlwein O, Robinson MJ, Dustan S et al (2011) DNA extraction columns contaminated with murine sequences. PLoS One 6:e23484CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Genetics, Institute for Integrative Systems BiologyUniversitat de ValènciaPaternaSpain

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