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. 2020 Sep;58(1):e110.
doi: 10.1002/cpmc.110.

The Propagation, Quantification, and Storage of Vesicular Stomatitis Virus

Alaa A Abdelmageed  1 , Maureen C Ferran  1
Affiliations

The Propagation, Quantification, and Storage of Vesicular Stomatitis Virus

Alaa A Abdelmageed et al. Curr Protoc Microbiol. 2020 Sep.

Abstract

Vesicular stomatitis virus (VSV) is the prototypical member of the Rhabdoviridae family of negative-sense single-stranded RNA viruses. This virus has been used as a powerful model system for decades and is currently being used as a vaccine platform and an oncolytic agent. Here, we present methods to propagate, quantitate, and store VSV. We also review the proper safety protocol for the handling of VSV, which is classified as a Biosafety Level 2 pathogen by the United States Centers for Disease Control and Prevention. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Generation, purification, and storage of vesicular stomatitis virus stocks Basic Protocol 2: Quantification of vesicular stomatitis virus by plaque assay Support Protocol: Propagation of Vero cells.

Keywords: plaque assay; propagation; quantification; stock; storage; titration; vesicular stomatitis virus.

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Conflict of interest statement

Conflict of Interest

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Plaque assay 6-well plate setup.
Figure 2:
Figure 2:
Image of a plaque assay showing the dilutions giving rise to different numbers of plaques. Note: These plaques are large because they were allowed to develop for 3 days to ensure that they were visible in the figure.
See this image and copyright information in PMC

References

    1. Agbulos DS, Barelli L, Giordano BV, & Hunter FF (2016). Zika Virus: Quantification, Propagation, Detection, and Storage. Curr Protoc Microbiol, 43, 15D.14.11–15D.14.16. doi:10.1002/cpmc.19 - DOI - PubMed
    1. Baer A, & Kehn-Hall K (2014). Viral concentration determination through plaque assays: using traditional and novel overlay systems. J Vis Exp(93), e52065. doi:10.3791/52065 - DOI - PMC - PubMed
    1. Fernandez M, Porosnicu M, Markovic D, & Barber GN (2002). Genetically engineered vesicular stomatitis virus in gene therapy: application for treatment of malignant disease. J Virol, 76(2), 895–904. doi:10.1128/jvi.76.2.895-904.2002 - DOI - PMC - PubMed
    1. Gélinas JF, Kiesslich S, Gilbert R, & Kamen AA (2020). Titration methods for rVSV-based vaccine manufacturing. MethodsX, 7, 100806. doi:10.1016/j.mex.2020.100806 - DOI - PMC - PubMed
    1. Hastie E, Besmer DM, Shah NR, Murphy AM, Moerdyk-Schauwecker M, Molestina C, . . . Grdzelishvili VZ (2013). Oncolytic vesicular stomatitis virus in an immunocompetent model of MUC1-positive or MUC1-null pancreatic ductal adenocarcinoma. J Virol, 87(18), 10283–10294. doi:10.1128/JVI.01412-13 - DOI - PMC - PubMed

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