Study for establishing differential diagnostic protocols for Japanese encephalitis virus and West Nile virus infection

• Project/Area Number: 18580301
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied veterinary science
• Research Institution: Hokkaido University
• Principal Investigator: MAEDA Akihiko Hokkaido University, Graduate school of Veterinary Science, Associate Professor (70333359)
• Co-Investigator(Kenkyū-buntansha): MAEDA Junko Hokkaido University, Graduate school of Veterinary Science, Technician (70399989)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,720,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥420,000); Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: Javanese encephalitis virus (JEV) / West Nile virus (WNV) / Differential diagnosis / Subviral particles (SvPs) / Virus-like particles (VLPs) / Replicon / Flaviviruses / Serological diagnosis / ELISA / ウエスタンブロット / 血清診断

Research Abstract

In this research program, we established the differential diagnostic protocols for flaviviruses, especially Japanese encephalitis virus (JEV) and West Nile virus (WNV). Because, JEV, which belongs to the same flavivirus serological group, in which WNV belongs, circulates in the nature of Japan, and the symptoms caused by JEV and WNV infections are quite similar, we should differentiate both infections to confirm the impost of WNV into Japan where is a virgin country of WNV infection. (1) We established the flavivirus reverse genetic systems, to produce subviral particles (SvPs), viral replicons and virus-like particles (VLPs). SvPs are the particle structures composed by only viral envelope proteins without viral related RNAs. The viral replicons are the RNAs, which derived from viral defective genomic RNAs and auto-replicate in the replicon-transfected cells. The VLPs are the particles composed of viral envelope proteins as its viral shells, and viral replicons within the shells. (2) We established differential diagnostic protocols, ELISA, IFA and Western blot analysis to differentiate JEV and WNV using SvPs of JEV and WNV. (3) We generated reporter protein-expressing WNV replies. VLPs composed. of JEV or WNV structural proteins as surface envelope proteins, and WNV replicons within the shells, were also generated. We are now establishing a modified-neutralizing test for differential diagnosis of flaviviruses using the VLPs.

Research Products

• [Journal Article] Detection of small RNAs containing the 5'-and the 3'-end sequences of viral genome during West Nile virus replication. (2008)
  • Author(s): Maeda, A., Maeda, J., Takagi, H., and Kurane, I.
  • Journal Title: Virology 371 Pages: 130-138
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A PCR-based protocol for generating West Nile virus replicons. (2008)
  • Author(s): Maeda, J., Takagi, H., Hashimoto, S., Kurane, I., and Maeda, A.
  • Journal Title: Journal of Virological Methods 148 Pages: 244-252
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Molecular characterization of the E gene of dengue virus type 1 isolated in Guangdong Province, China, in 2006. (2008)
  • Author(s): Zheng, K., Zhou, H.-Q., Yan, J., Ke, C.-W., Maeda, A., Maeda, J., Takashima, I., Kurane, I., Ma, H., and Xie, X.-M.
  • Journal Title: Epidemiology and Infection (印刷中)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Detection of small RNAs containing the 5'- and the 3'-end sequences of viral genome during West Nile virus replication. (2008)
  • Author(s): Maeda A., Maeda, J., Takagi, H., and Kurane, I.
  • Journal Title: Virology 371 Pages: 130-138
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A PCR-based protocol for generating West Nile virus replicons. (2008)
  • Author(s): Maeda, J., Takagi, H., Hashimoto, S., Kurane, I., and Maeda. A.
  • Journal Title: Journal of Virological Methods 148 Pages: 244-252
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular characterization of the E gene of dengue virus type 1 isolated in Guangdong Province, China, in 2006. (2008)
  • Author(s): Zheng, K., Zhou, H.-Q., Yan, J., Ke, C.-W. Maeda, A., Maeda, J., Takashima, I., Kurane, I., Ma, H., and Xie, X. -M.
  • Journal Title: Epidemiology and Infection (In press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Detection of small RNAs containing the 5'-and the 3'-end sequences of viral genome during West Nile virus replication. (2008)
  • Author(s): maeda, A., Maeda, J., Takagi, H., and Kurane, I.
  • Journal Title: Virology 371 Pages: 130-138
  • Peer Reviewed
• [Journal Article] 輸入牧草に混入し北海道にてその死体が確認されたホシムクドリSturnus vulgarisの記録とウイルス学的検査 (2007)
  • Author(s): 吉野智生、国藤泰輔、渡辺竜己、久木田優美、前田秋彦、荻原克郎、大沼学、桑名貴、村田浩一、浅川満彦
  • Journal Title: 北海道獣医師会雑誌 51 Pages: 68-70
  • NAID: 120005761172
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Virological test on Sturnus vulgaris, which was contaminated in imported glasses to Hokkaido. (2007)
  • Author(s): Yoshino, I., Kunifuji, Y., Kubota, Y., Maeda, A., Ogiwara, K., Oonuma, M., Kuwana, T., Murata, K., and Asanaka, M.
  • Journal Title: Hokkaido Jyuuishi-kai Zasshi 51 Pages: 68-70
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 輸入牧草に混入し北海道にてその死体が確認されたホシムクドリSturnus vulgarisの記録とウイルス学的検査 (2007)
  • Author(s): 吉野智生、国藤泰輔、渡辺竜己、久木田優美、前田秋彦、荻原克郎、大沼学、桑名貴、村田浩一、浅川満彦
  • Journal Title: 北海道獣医師会雑誌 57 Suppl Pages: 46-46
  • NAID: 120005761172
• [Journal Article] 輸入牧草に混入し北海道にてその死体が確認されたホシムクドリSturnus vulgarisの記録とウイルス学的検査. (2007)
  • Author(s): 吉野智生, 国藤泰輔, 渡邊竜巳, 久保木優美, 前田秋彦, 荻原克郎, 大沼学, 桑名貴, 村田浩一, 浅川満彦
  • Journal Title: 北獣会誌 51 Pages: 68-70
  • NAID: 120005761172
• [Journal Article] Epidemiological study of mosquitoes borne-Flaviviruses in Guandong, China, 2005. (2006)
  • Author(s): Maeda, J., Ma, H., Sun, H., Ke, C.-G., Takagi, H., Kurane, I., Horiuchi, M., Ikuo, T., and Maeda, A.
  • Journal Title: Medical Entomology and Zoology 57 Suppl Pages: 46-46
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Epidemiological study of mosquitoes borne- Flaviviruses in Guandong, China, 2005. (2006)
  • Author(s): Maeda, J., Ma, H., Sun, H., Ke, C.-G., Takagi, H., Kurane, I., Horiuchi, M., Ikuo, T., and Maeda, A.
  • Journal Title: Medical Entomology and Zoology 57 Suppl Pages: 46-46
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] ウエストナイルウイルス E 蛋白質糖鎖付加のウイルス粒子形成と細胞変性効果への影響 (2007)
  • Author(s): 前田秋彦、前田潤子、橋本新吾、高木弘隆、高島郁夫、倉根一郎
  • Organizer: 第42回日本脳炎ウイルス生態学研究会
  • Place of Presentation: 石川、金沢医科大学
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 酪農学園大学野生動物医学センターWAMCにおける野鳥病原体感染のリスク評価研究(概要紹介) (2007)
  • Author(s): 浅川満彦、大沼学、吉野智生、佐々木均、前田秋彦、長嶺隆、天野洋祐、伊東孝、斉藤美加、外平友佳理、村田浩一、桑名貴
  • Organizer: 第13回日本野生動物医学会
  • Place of Presentation: 岩手、いわて県民交流センター
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 中空ウイルス粒子を用いたウエストナイルと日本脳炎ウイルスに対する感染血清の鑑別 (2007)
  • Author(s): 前田潤子、高木弘隆、倉根一郎、高島郁夫、前田秋彦
  • Organizer: 第144回日本獣医学会
  • Place of Presentation: 江別、酪農学園大学
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 2006年、中国・広東省で分離されたデングウイルスの分子疫学 (2007)
  • Author(s): 前田秋彦、前田潤子、高木弘隆、高島郁夫、倉根一郎
  • Organizer: 第55回日本ウイルス学会
  • Place of Presentation: 札幌、札幌コンベンションセンター
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Effects of glycosylation of West Nile virus E proteins on viral assembly and cytopathological effects (2007)
  • Author(s): Maeda A., Maeda, J., Hashimoto, S., Takagi, H., Takashima, I., Kurane, I.
  • Organizer: 42^<th> Annual meeting of The Society of Ecology on Japanese encephalitis virus
  • Place of Presentation: Ishikawa, Medical University of Kanazawa
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Study of risk analysis on birds infectious diseases in Wild Animal Medical Center (WAMC) of Rakuno Gakuen University (Over view) (2007)
  • Author(s): Asakawa, M., Oonuma, M., Yoshino, T., Sasaki, H., Maeda, A., Nagamine, T., Amano, Y., Itoh, T., Saito, M., Tohira, Y, Murata, K., and Kuwana, T.
  • Organizer: 13^<th> Annual meeting of The Japanese Society of Zoo and Wildlife Medicine
  • Place of Presentation: Iwate, Iwate Prefecture Conventional Center
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Differential diagnosis of West Nile virus and Japanese encephalitis virus using Subviral particles (2007)
  • Author(s): Maeda, J., Takagi, H., Kurane, I., Takashima, I., Kurane, I., Maeda, A.
  • Organizer: 144^<th> Annual meeting of The Japanese Society of Veterinaly Science
  • Place of Presentation: Ebetu, Rakuno Gakuen University
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Molecular epidemiological analysis on dengue virus outbreak in 2006, in Guangdong, China (2007)
  • Author(s): Maeda, A., Maeda, J., Takagi, H., Takashima, I., Kurane, I.
  • Organizer: 55^<th> Annual meeting of The Japanese Society for Virology
  • Place of Presentation: Sapporo, Sapporo Conventional Center
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] ウエストナイルウイルスE蛋白質糖鎖付加のウイルス粒子形成と細胞変性効果への影響 (2007)
  • Author(s): 前田秋彦、前田潤子、橋本新吾、高木弘隆、高島郁夫、倉根一郎
  • Organizer: 第42回日本脳炎ウイルス生態学研究会
  • Place of Presentation: 石川、金沢医科大学
• [Presentation] 酪農学園大学野生動物医学センターWAMCにおける野鳥病原体感染のリスク評価研究(概要紹介) (2007)
  • Author(s): 浅川満彦、大沼学、吉野智生、佐々木均、前田秋彦、長嶺隆、天野洋祐、伊東孝、斉藤美加、外平友佳理、村田浩一、桑名貴
  • Organizer: 第13回日本野生動物医学会
  • Place of Presentation: 岩手、いわて県民情報交流センター
• [Presentation] 地方衛研におけるバイオセーフティ研修実施の試み(第2報) (2006)
  • Author(s): 伊木繁雄、高木弘隆、後藤浩、川又亨、地主勝、森本洋、池田徹也、駒込理佳、木村浩一、清水俊一、山口敬治、前田秋彦、杉山和良、岡野素彦
  • Organizer: 第6回日本バイオセーフティー学会
  • Place of Presentation: 横浜、明治製菓本社講堂
  • Year and Date: 2006-11-24
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] The attempt of the biosafety training in Hokkaido Institute of Public Health (2^<nd> report) (2006)
  • Author(s): Igi, S., Takagi, H., Goto, H., Kawamata, T., Zinushi, M., Morimoto, T., Komagome, R., Kimura, H., Shimizu, S., Yamaguchi, K., Maeda, A., Sugiyama, K., and Okano, M.
  • Organizer: 6^<th> Annual meeting of The Japanese Biological Safety Association
  • Place of Presentation: Yokohama, Meiji-Seika
  • Year and Date: 2006-11-24
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 2005年、中国・広東省珠海市で採取した蚊のフラビウイルス保有状況の調査 (2006)
  • Author(s): 前田潤子、馬洪波、孫虹、柯昌文, 高木弘隆、倉根一郎、堀内基広、高島郁夫、前田秋彦
  • Organizer: 第58回日本衛生動物学会
  • Place of Presentation: 長崎、長崎大学
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] アジアにおける蚊媒介性感染症の疫学的解析-中国を中心に- (2006)
  • Author(s): 前田秋彦、Ma Hongbo、 Hong Sun、 Ke Chuang-Wen、高木弘隆、前田潤子、堀内基広、高島郁夫、倉根一郎
  • Organizer: 第41回日本脳炎ウイルス生態学研究会
  • Place of Presentation: 長崎、長崎大学
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ウエストナイルウイルスのレプリコンの作製 (2006)
  • Author(s): 橋本新吾、前田潤子, 高木弘隆、堀内基広、高島郁夫、倉根一郎、前田秋彦
  • Organizer: 第54回日本ウイルス学会
  • Place of Presentation: 名古屋、名古屋国際会議場
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ウエストナイルウイルスのマイナス鎖smallRNAの合成 (2006)
  • Author(s): 前田秋彦、前田潤子、高木弘隆、堀内基広、倉根一郎
  • Organizer: 第54回日本ウイルス学会
  • Place of Presentation: 名古屋、名古屋国際会議場
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Epidemiological study on mosquito- borne borne flaviviruses, in Guangdong, China, in 2005 (2006)
  • Author(s): Maeda, J., Ma, K., Sun, H., Ke, C-. W., Takagi, H., Kurane, I., Horiuchi, M., Takashima, I., and Maeda, A.
  • Organizer: 58^<th> Annual meeting of The Japan Society of Medical Entomology and Zoology
  • Place of Presentation: Nagasaki, Nagasaki University
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Epidemiological study mosquito-borne flaviviruses, in Asia (2006)
  • Author(s): Maeda, A., Ma, H., Sun, H., Ke, C-. W., Takagi, H., Maeda, J., Horiuchi, M., Takashima, I., Kurane, I.
  • Organizer: 41^<th> Annual meeting of The Society of Ecology on Japanese encephalitis virus
  • Place of Presentation: Nagasaki, Nagasaki University
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Synthesis of West Nile virus Replicon (2006)
  • Author(s): Hashimoto, S., Maeda, J., Takagi, H., Horiuchi, M., Takashima, I., Kurane, I., and Maeda, A.
  • Organizer: 54^<th> Annual meeting of The Japanese Society for Virology
  • Place of Presentation: Nagoya, Nagoya International Center
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Detection of West Nile virus minus-strand RNAs in infected cells. (2006)
  • Author(s): Maeda, A., Maeda, J., Takagi, H., Horiuchi, M., Kurane, I.
  • Organizer: 54^<th> Annual meeting of The Japanese Society for Virology
  • Place of Presentation: Nagoya, Nagoya International Center
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] SUMOylation viral infection: Dose SUMO erestle or help for viral infection? In Current Topics in Biochemistry (2006)
  • Author(s): Maeda, A., Maeda, J., and I., Kurane
  • Total Pages: 150
  • Publisher: Research Signpost, Kerala, India
  • Description: 「研究成果報告書概要(和文)」より
• [Book] SUMOylation in viral infection: Dose SUMO wrestle or help for viral infection? (2006)
  • Author(s): Maeda, A., Maeda, J., and Kurane, I.
  • Publisher: In Current Topics in Biochemistry, Ed. Ohyama, T., Research Signpost, Kerala, India
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] Current topics in Biochemistry (SUMOylation in viral infection : Dose SUMO wressle against or help for viral infection.) (2006)
  • Author(s): Maeda, A., Maeda, J., Kurane, I.
  • Total Pages: 150
  • Publisher: RESEARCH SIGNPOST