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Article
PCR-Activated Cell Sorting as a General, Cultivation-Free Method for High-Throughput Identification and Enrichment of Virus Hosts
Journal of Virological Methods
  • Shaun W. Lim, University of California, San Francisco
  • Shea T. Lance, University of California, San Francisco
  • Kenneth M. Stedman, Portland State University
  • Adam R. Abate, University of California, San Francisco
Document Type
Post-Print
Publication Date
4-1-2017
Subjects
  • Host-virus relationships,
  • Bacteria -- Identification,
  • Polymerase chain reaction,
  • Molecular biology -- Technique
Disciplines
Abstract

Characterizing virus-host relationships is critical for understanding the impact of a virus on an ecosystem, but is challenging with existing techniques, particularly for uncultivable species. We present a general, cultivation-free approach for identifying phage-associated bacterial cells. Using PCR-activated cell sorting, we interrogate millions of individual bacteria for the presence of specific phage nucleic acids. If the nucleic acids are present, the bacteria are recovered via sorting and their genomes analyzed. This allows targeted recovery of all possible host species in a diverse population associated with a specific phage, and can be easily targeted to identify the hosts of different phages by modifying the PCR primers used for detection. Moreover, this technique allows quantification of free phage particles, as benchmarked against the “gold standard” of virus enumeration, the plaque assay.

Description

This is the author’s version of a work that was accepted for publication in Journal of Virological Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Virological Methods, http://dx.doi.org/10.1016/j.jviromet.2016.12.009

© 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/

DOI
10.1016/j.jviromet.2016.12.009
Persistent Identifier
http://archives.pdx.edu/ds/psu/18934
Citation Information
Lim, ShaunW., Lance, Shea T., Stedman, Kenneth M., Abate, Adam R., PCR-Activated Cell Sorting as a General, Cultivation-Free Method for High-Throughput Identification and Enrichment of Virus Hosts. Journal of Virological Methods http://dx.doi.org/10.1016/j.jviromet.2016.12.009