Next generation sequencing shows West Nile virus quasispecies diversification after a single passage in a carrion crow (Corvus corone) in vivo infection model

Dridi, M., Lecollinet, S., Orton, R. , Johnson, P. , Rosseel, T., van Borm, S., Lambrecht, B. and Muylkens, B. (2015) Next generation sequencing shows West Nile virus quasispecies diversification after a single passage in a carrion crow (Corvus corone) in vivo infection model. Journal of General Virology, 96, p. 2999. (doi:10.1099/jgv.0.000231) (PMID:26297666)

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Abstract

West Nile virus (WNV) occurs as a population of genetic variants (quasispecies) infecting a single animal. Previous low resolution viral genetic diversity estimates in sampled wild birds and mosquitoes and in multiple passage adaptation studies in vivo or in cell culture, suggest that WNV genetic diversification is mostly limited to the mosquito vector. This study investigates genetic diversification of WNV in avian hosts during a single passage using next generation sequencing. Wild-captured Carrion crows were subcutaneously infected using a clonal Middle-East WNV. Blood samples were collected on 2 and 4 days post-infection. A RT-PCR approach was used to amplify the WNV genome directly from serum samples prior to next generation sequencing resulting in an average depth of at least 700x in each sample. Appropriate controls were sequenced to discriminate biologically relevant low frequency variants from experimentally introduced errors. The WNV populations in the wild crows showed significant diversification away from the inoculum virus quasispecies structure. By contrast, WNV populations in intracerebrally infected day-old chickens did not diversify from that of the inoculum. Where previous studies concluded that WNV genetic diversification is only experimentally demonstrated in its permissive insect vector species, we have experimentally shown significant diversification of WNV populations in a wild bird reservoir species.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Johnson, Dr Paul and Orton, Dr Richard
Authors: Dridi, M., Lecollinet, S., Orton, R., Johnson, P., Rosseel, T., van Borm, S., Lambrecht, B., and Muylkens, B.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Journal of General Virology
Publisher:Society for General Microbiology
ISSN:0022-1317
ISSN (Online):1465-2099
First Published:First published in Journal of General Virology 96:2999-3009
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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