Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse

James D. Brien, Mariah Hassert, E. Taylor Stone, Elizabeth Geerling, Lillian Cruz-Orengo, Amelia K. Pinto

Research output: Contribution to journalArticle

Abstract

The methods being presented demonstrate laboratory procedures for the isolation of organs from Zika virus infected animals and the quantification of viral load. The purpose of the procedure is to quantify viral titers in peripheral and CNS areas of the mouse at different time points post infection or under different experimental conditions to identify virologic and immunological factors that regulate Zika virus infection. The organ isolation procedures demonstrated allow for both focus forming assay quantification and quantitative PCR assessment of viral titers. The rapid organ isolation techniques are designed for the preservation of virus titer. Viral titer quantification by focus forming assay allows for the rapid throughput assessment of Zika virus. The benefit of the focus forming assay is the assessment of infectious virus, the limitation of this assay is the potential for organ toxicity reducing the limit of detection. Viral titer assessment is combined with quantitative PCR, and using a recombinant RNA copy control viral genome copy number within the organ is assessed with low limit of detection. Overall these techniques provide an accurate rapid high throughput method for the analysis of Zika viral titers in the periphery and CNS of Zika virus infected animals and can be applied to the assessment of viral titers in the organs of animals infected with most pathogens, including Dengue virus.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number150
DOIs
StatePublished - Aug 15 2019

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Viruses
Assays
Viral Load
Limit of Detection
Animals
Animal Structures
Polymerase Chain Reaction
Dengue Virus
Viral Genome
Immunologic Factors
Throughput
Pathogens
Zika Virus
RNA
Toxicity
Genes
Infection

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse. / Brien, James D.; Hassert, Mariah; Stone, E. Taylor; Geerling, Elizabeth; Cruz-Orengo, Lillian; Pinto, Amelia K.

In: Journal of visualized experiments : JoVE, No. 150, 15.08.2019.

Research output: Contribution to journalArticle

Brien, James D. ; Hassert, Mariah ; Stone, E. Taylor ; Geerling, Elizabeth ; Cruz-Orengo, Lillian ; Pinto, Amelia K. / Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse. In: Journal of visualized experiments : JoVE. 2019 ; No. 150.
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