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Research Project: Rift Valley Fever Pathogenesis, Epidemiology, and Control Measures

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Title: Complete genome sequence of a 2016 Bluetongue virus serotype 3 isolate from Louisiana, USA

Author
item Schirtzinger, Erin
item RUDER, MARK - University Of Georgia
item STALLKNECHT, DAVID - University Of Georgia
item Wilson, William

Submitted to: Microbiology Resource Announcements
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/16/2019
Publication Date: 5/9/2019
Citation: Schirtzinger, E.E., Ruder, M.G., Stallknecht, D.E., Wilson, W.C. 2019. Complete genome sequence of a 2016 Bluetongue virus serotype 3 isolate from Louisiana, USA. Microbiology Resource Announcements. 8:e00128-19. https://doi.org/10.1128/MRA.00128-19.
DOI: https://doi.org/10.1128/MRA.00128-19

Interpretive Summary: The full-genome sequence of BTV-3 USA2016/LA CC16-564 isolated from a white-tailed deer in East Feliciana Parish, Louisiana is reported here. Nine genomic segments of this virus have 99% identity with a 2013 BTV-3 isolate from Florida, while segment 10 has 97% identity with a 2003 BTV-5 isolate from Florida.

Technical Abstract: BTV-3 USA2016/LA CC16-564 was isolated from a white-tailed deer in East Feliciana Parish, Louisiana in 2016. The virus was passaged twice in baby hamster kidney cells (BHK-21) to produce sufficient virus for full-genome sequencing. Double-stranded RNA was isolated from total RNA by lithium chloride differential separation and subjected to whole genome sequencing using the sequence–independent amplification procedure. The library was prepared with the Nextera XT DNA Library Prep Kit and Nextera XT Index Kit (Illumina) according to manufacturer’s instructions. The library was normalized manually and sequenced on the Illumina MiSeq. Paired-end reads were trimmed of low quality bases and de novo assembled into contigs using CLC Genomics Workbench 11.0 (Qiagen). Contigs were compared to the GenBank nucleotide database using the BLAST function to determine a suitable reference for reference-based assembly. Phylogenetic analyses of each of the BTV-3 genome segments with those of other US, Central American and Caribbean BTVs have shown that nine genomic segments of this virus have 99% identity with a 2013 BTV-3 isolate from Florida, while segment 10 has 97% identity with a 2003 BTV-5 isolate from Florida.