Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing

Rapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical propert...

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Main Authors: Mya Breitbart, Forest Rohwer
Format: Article
Language:English
Published: Taylor & Francis Group 2005-11-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/000112019
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author Mya Breitbart
Forest Rohwer
author_facet Mya Breitbart
Forest Rohwer
author_sort Mya Breitbart
collection DOAJ
description Rapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical properties of viruses combined with sequence-independent amplification and cloning. We show that both single- and double-stranded DNA viruses can be recovered from blood samples using this approach. In addition, we report the discovery of novel anellovirus sequences in the blood of healthy donors. PCR primers designed to amplify these novel anellovirus sequences were then used to verify the presence of these viruses in the general donor population.
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1940-9818
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spelling doaj-art-4d421b1f8e65401b9195aba931e76ba72025-08-20T02:25:37ZengTaylor & Francis GroupBioTechniques0736-62051940-98182005-11-0139572973610.2144/000112019Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencingMya Breitbart0Forest Rohwer11San Diego State University, San Diego, CA, USA1San Diego State University, San Diego, CA, USARapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical properties of viruses combined with sequence-independent amplification and cloning. We show that both single- and double-stranded DNA viruses can be recovered from blood samples using this approach. In addition, we report the discovery of novel anellovirus sequences in the blood of healthy donors. PCR primers designed to amplify these novel anellovirus sequences were then used to verify the presence of these viruses in the general donor population.https://www.future-science.com/doi/10.2144/000112019
spellingShingle Mya Breitbart
Forest Rohwer
Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
BioTechniques
title Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
title_full Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
title_fullStr Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
title_full_unstemmed Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
title_short Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing
title_sort method for discovering novel dna viruses in blood using viral particle selection and shotgun sequencing
url https://www.future-science.com/doi/10.2144/000112019
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