Abstract
In this study, we improved a method for rapid determination of viral RNA sequences (RDV) to overcome the limitations of previous versions. The RDV ver4.0 method can detect RNA sequences with at least 1,000 copies as starting material. A novel virus, which was isolated from field-collected Aedes aegypti larvae in the Phasi Charoen district of Thailand using C6/36 cells, was identified using the RDV ver4.0 protocol. The virus was named Phasi Charoen virus (PhaV). We used a high-throughput pyrosequencing approach to obtain more information about the genome sequence of PhaV. Analysis of a phylogenic tree based on amino acid sequences strongly suggested that PhaV belongs to the family Bunyaviridae.
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Acknowledgments
We thank Ms. Momoko Ogata, Itoe Iizuka, Tomoyuki Shiota of the National Institute of Infectious Diseases of Japan, for her assistance, and also Ms. Parichat Lapcharoene, Pornlada Nuchnoi, Somruay Srumnam, Jittarat Srithong and Aisa Togvang, of the Faculty of Tropical Medicine, Mahidol University for their mosquito collection. This study was supported in part by a Grant from The Japan Society for the Promotion of Science, by the Ministry of Health, Labor, and Welfare, and by the Ministry of Education, Culture, Sports, Science and Technology, Japan.
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Yamao, T., Eshita, Y., Kihara, Y. et al. Novel virus discovery in field-collected mosquito larvae using an improved system for rapid determination of viral RNA sequences (RDV ver4.0). Arch Virol 154, 153–158 (2009). https://doi.org/10.1007/s00705-008-0285-5
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DOI: https://doi.org/10.1007/s00705-008-0285-5