Laboratory preparation of Varicella-Zoster Virus: Concentration of virus-containing supernatant, use of a debris fraction and magnetofection for consistent cell-free VZV infections

Anna Sloutskin, Ronald S. Goldstein

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

The research laboratory generation of free Varicella-Zoster Virus (VZV) from cultured yields results relatively low titers, with the result that most study of VZV infection utilizes cell-associated infection. However, important aspects of VZV-cell interaction, such as the entry mechanism and superinfection exclusion have not yet been studied in detail, in part due to the difficulty in obtaining a high titer cell free virus. Here, a method to generate relatively high-titer cell-free VZV, based on a combination of previously published techniques and subsequent concentration is described. VZV-infected cells are disrupted, sonicated and clarified by centrifugation. The cell-free virus in the supernatant is then concentrated to yield up to 105PFU/ml. The cell debris pellet, which contains up to 106PFU/ml can also be used for non cell-associated infection. Magnetic nanoparticles available commercially can be used to further enhance infection by cell-free-VZV. The tools described here hold promise for better understanding of important aspects of VZV-cell interactions such as entry and latency.

Original languageEnglish
Pages (from-to)128-132
Number of pages5
JournalJournal of Virological Methods
Volume206
DOIs
StatePublished - 15 Sep 2014

Bibliographical note

Funding Information:
This study was supported by Israel Science Foundation grant #238/11 and a generous gift from the Maximillian Goode Foundation to RSG. We thank Prof. Paul R. Kinchington (University of Pittsburgh, Pittsburgh PA) for the generous gift of fluorescent viruses, as well as his expert advice and support of the study. Thanks as always to Chaya Morgenstern for expert technical and logistic support.

Funding

This study was supported by Israel Science Foundation grant #238/11 and a generous gift from the Maximillian Goode Foundation to RSG. We thank Prof. Paul R. Kinchington (University of Pittsburgh, Pittsburgh PA) for the generous gift of fluorescent viruses, as well as his expert advice and support of the study. Thanks as always to Chaya Morgenstern for expert technical and logistic support.

FundersFunder number
Maximillian Goode Foundation
Israel Science Foundation238/11

    Keywords

    • Cell-free virus
    • Magnetic infection
    • Varicella-Zoster Virus

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