Objective To investigate the diversity of mosquito-borne viruses in Xinjiang , China, and to identify mosquitos-borne viruses of medical importance rapidly .Methods The virus-derived RNAs in mosquitos captured in wild were screened and confirmed by using Illumina deep sequencing approach and reverse transcription PCR , respectively .The alignment analysis was performed by using gene sequences from GenBank.Results One hundred and forty-four Culex Flavivirus ( CxFV, Flavivirus genus, Flaviviridae) specific sequences were identified .The overlapping reads were assembled into 7 uncontinuous viral genomic contigs.The gaps between the contigs were further filled by RT-PCR products, which resulted in reconstruc-tion of viral genomic 5′and 3′terminus (687 nt and 411 nt).Phylogenetic analysis showed that the newly identified CxFV belonged to America/Asian genotype , which specifically clustered into a clade with other CxFV strains from China mainland ,sharing 98.2%-99.5%homologies in nucleotide sequences and 99.5%in amino acids sequences among them .Conclusion Illumina deep sequencing approach was successfully applied to arthropod-borne virus surveillance .The recently emerged Culex Flavivirus was detected for the first time in Xinjiang, China.

OBJECTIVETo monitor and discover medically important mosquito-borne viruses circulating in Xinjiang, China.

METHODSMosquitoes were collected from Abbey Lake wetland in Bortala, in Northern Xinjiang. Viral isolates were obtained through inoculating and serial passaging into susceptible mammalian host cells (BHK-21), identified by cytopathogenic effect (CPE) observation and plague forming assay. Genetic identification of viral isolates was conducted by RT-PCR, sequencing and phylogenetic analysis.

RESULTSA virus strain which causing CPE on BHK-21 cells, was isolated from the predominant Culex modestus (36.6%) and tentatively designated as Abbey Lake virus. Information on molecular identification revealed that Abbey Lake virus belonged to Orthobunyavirus genus within Bunyaviridae. Partial sequences (651 bp and 980 bp) of viral genomic S and M segment showed that Abbey Lake virus was phylogenetically related to Germiston virus that uniquely found in South Africa with 90.6% nucleotides and 95.0% amino acids similarities in S segment. However, viral M segment displayed much variability with 78.6% nucleotides and 86.1% amino acid similarities, suggesting a new member of Orthobunyavirus genus was discovered in the area.

CONCLUSIONIn this study, Abbey Lake virus was isolated and characterized indicating its potential circulation nature of this newly-emerged mosquito-borne virus.

Animals ; China ; Culicidae ; virology ; Orthobunyavirus ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Analysis, RNA ; Viral Proteins ; genetics