Abstract
We describe the isolation and characterization of a novel flavivirus, isolated from a pool of Culex (Melanoconion) ocossa Dyar and Knab mosquitoes collected in 2009 in an urban area of the Amazon basin city of Iquitos, Peru. Flavivirus infection was detected by indirect immunofluorescent assay of inoculated C6/36 cells using polyclonal flavivirus antibodies (St. Louis encephalitis virus, yellow fever virus and dengue virus type 1) and confirmed by RT-PCR. Based on partial sequencing of the E and NS5 gene regions, the virus isolate was most closely related to the mosquito-borne flaviviruses but divergent from known species, with less than 45 and 71 % pairwise amino acid identity in the E and NS5 gene products, respectively. Phylogenetic analysis of E and NS5 amino acid sequences demonstrated that this flavivirus grouped with mosquito-borne flaviviruses, forming a clade with Nounané virus (NOUV). Like NOUV, no replication was detected in a variety of mammalian cells (Vero-76, Vero-E6, BHK, LLCMK, MDCK, A549 and RD) or in intracerebrally inoculated newborn mice. We tentatively designate this genetically distinct flavivirus as representing a novel species, Nanay virus, after the river near where it was first detected.
Highlights
The genus Flavivirus comprises, at the time of writing, approximately 70 virus species, there are probably many more species yet undiscovered (Pybus et al, 2002)
Attempts to culture the virus in mammalian cells, including simian (LLCMK, Vero-76 and Vero-E6), hamster (BHK), canine (MDCK) and human (RD and A549) cells were unsuccessful, as no cytopathic effects were observed and all cultures were negative by immunofluorescence assay (IFA) and RT-PCR after multiple passages
We report the identification and characterization of a novel flavivirus isolated from Culex (Melanoconion) ocossa Dyar and Knab mosquitoes from Iquitos, Peru
Summary
The genus Flavivirus (family Flaviviridae) comprises, at the time of writing, approximately 70 virus species, there are probably many more species yet undiscovered (Pybus et al, 2002). Mosquito-borne flaviviruses and tick-borne flaviviruses, including yellow fever virus (YFV), dengue virus serotypes 1–4 (DENV-1 to DENV-4), Japanese encephalitis virus (JEV), West Nile virus (WNV) and tick-borne encephalitis virus (TBEV), are often associated with human infections and are the cause of high morbidity and mortality worldwide. Flaviviruses such as Culex flavivirus (CxFV), Kamiti River virus (KRV) and cell fusing agent virus (CFAV) have only been isolated from mosquitoes or mosquito cell lines, have no recognized association with vertebrates, and have not yet been associated with human or animal disease. While not responsible for human disease, these viruses are of increasing interest owing to possible interactions in the mosquito host with
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