Full-length genome sequence of Ntaya virus
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Presentation of pyrosequencing data and phylogenetic analysis for the full genome of Ntaya virus, type virus of the Ntaya virus group of the Flaviviridae isolated in Cameroon in 1966.
KeywordsNtaya virus Flavivirus 454 Pyrosequencing
The Flavivirus Ntaya virus (NTAV) was originally isolated from mosquitoes in Uganda in 1951  and serologically determined as the type virus of the Ntaya virus group within the Flaviviridae . Serosurveys have detected NTAV activity in migratory birds and domestic animals in Romania [3, 4, 5, 6]. Antibodies against NTAV in travellers from Africa indicate transmission in Uganda, Cameroon, Democratic Republic of Congo, Kenya, Nigeria and Zambia. Clinical evidence indicated neurological manifestations of disease .
A lyophilised 10 % suckling mouse brain suspension of NTAV isolate IPD/A of the CRORA collection at the Institute Pasteur Dakar collected in Cameroon in 1966 was passaged twice on Vero E6 cells in 175-cm2 tissue culture flasks (DMEM, 2 % FBS, 2 mM glutamine, 10 mM penicillin, 10 mM streptomycin and 20 mM HEPES) at 37 °C and 5 % CO2. At 90–100 % CPE (12 dpi), culture supernatants of infected cells were collected, and purification and RNA extraction were performed as described . In order to determine the termini, a self-complimentary 3′-FLAC adapter was ligated to the 3′ end and a 5′-RACE adapter was ligated to the 5′ end of the +ssRNA genome prior to pyrosequencing as described . The complete genome was determined in a pool of seven MID-tagged virus libraries in one pyrosequencing run. Bioinformatic analysis was performed as described . The genome size was 10,891 bp (GenBank JX236040). 93 % of 4,730 reads were specific for NTAV (coverage 116-fold). The NTAV genome was assembled by reference mapping and showed the highest nucleic acid identity (77 %) to strains of Bagaza virus (10,284–10,941 bp).
This work was supported by the Federal Ministry of Education and Research (BMBF), grant number 01KI0710, ‘Research on Zoonotic Infectious Diseases’ programme, ‘Emerging arthropode-borne viral infections in Germany: Pathogenesis, diagnostics and surveillance’ and the BMBF funded research programme ‘Potential release-oriented biothreat emergency diagnostics (P.R.O.B.E)’ for civil security of the German Federal Government as part of the high-tech strategy for Germany.
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