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Virology. 2018 Aug;521:138-148. doi: 10.1016/j.virol.2018.06.004. Epub 2018 Jun 20.

Using barcoded Zika virus to assess virus population structure in vitro and in Aedes aegypti mosquitoes.

Author information

1
Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United States. Electronic address: weger@vt.edu.
2
Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United States.
3
Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, WI, United States.
4
Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, WI, United States.
5
Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, WI, United States; Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, United States.
6
Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, WI, United States; Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, United States.
7
Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United States. Electronic address: gregory.ebel@colostate.edu.

Abstract

Arboviruses such as Zika virus (ZIKV, Flaviviridae; Flavivirus) must replicate in both mammalian and insect hosts possessing strong immune defenses. Accordingly, transmission between and replication within hosts involves genetic bottlenecks, during which viral population size and genetic diversity may be significantly reduced. To help quantify these bottlenecks and their effects, we constructed 4 "barcoded" ZIKV populations that theoretically contain thousands of barcodes each. After identifying the most diverse barcoded virus, we passaged this virus 3 times in 2 mammalian and mosquito cell lines and characterized the population using deep sequencing of the barcoded region of the genome. C6/36 maintain higher barcode diversity, even after 3 passages, than Vero. Additionally, field-caught mosquitoes exposed to the virus to assess bottlenecks in a natural host. A progressive reduction in barcode diversity occurred throughout systemic infection of these mosquitoes. Differences in bottlenecks during systemic spread were observed between different populations of Aedes aegypti.

KEYWORDS:

Arbovirus; Flavivirus; Mosquito-borne virus; Virus evolution; Zika virus

PMID:
29935423
PMCID:
PMC6309320
DOI:
10.1016/j.virol.2018.06.004
[Indexed for MEDLINE]
Free PMC Article

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