Development of method for comprehensive isolation and purification of viral particles from clinical specimens

• Project/Area Number: 23659298
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: NAKAYA Takaaki 京都府立医科大学, 大学院・医学研究科, 教授 (80271633)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2011: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
• Keywords: ウイルス / ウイルス粒子精製 / インフルエンザウイルス / ニューカッスル病ウイルス / メタゲノム

Research Abstract

A more effective method to purify viral paricles from clinical specimens is required in metagenomic analysis of pathogens using a high-throughput DNA sequencer. In this study, a novel virion purification method using HPLC or field flow fractionation was evaluated. Even in the preliminary results, field flow fractionation combined with a light scattering detector could isolate the virion fraction from a crude virus solution, such as influenza and Newcastle disease virus. In addition, it was demonstrated that hydroxyapatite chromatography effectively purified viral particles from allantoic fluid from an infected embryonated egg. These results suggested that the above methods could be useful for virion fractionation from crude samples.

Research Products

• [Journal Article] Human monoclonal antibodies broadly neutralizing against influenza B virus. (2013)
  • Author(s): Yasugi, M., Kubota-Koketsu, R., Yamashita, A., Kawashita, N., Du, A., Sasaki, T., Nishimura, M., Misaki, R., Kuhara, M., Boonsathorn, N., Fujiyama, K., Okuno, Y., Nakaya, T., and Ikuta, K.
  • Journal Title: PLoS Pathog. Volume: 9 Issue: 2 Pages: e1003150-e1003150
  • DOI: 10.1371/journal.ppat.1003150
  • Peer Reviewed
• [Journal Article] 次世代シークエンサーのウイルス性疾患同定への適用 (2012)
  • Author(s): 中屋隆明
  • Journal Title: 臨床とウイルス Volume: 40(No.4) Pages: 177-183
  • NAID: 10031123614
• [Journal Article] メタゲノム解析を応用した網羅的病原体検出法 (2012)
  • Author(s): 中村昇太, 中屋隆明, 飯田哲也
  • Journal Title: 最新医学 Volume: 67(No.9) Pages: 131-133
  • NAID: 40019431074
• [Journal Article] Emergence of HA mutants during influenza virus pneumonia (2012)
  • Author(s): Manriquez ME, Nakaya T, Shinya K, et al.
  • Journal Title: Int. J. Clin. Exp. Pathol. Volume: 5(8) Pages: 787-795
  • Peer Reviewed
• [Journal Article] Antigenic analysis of highly pathogenic avian influenza virus H5N1 sublineage co-circulating in Egypt. (2012)
  • Author(s): Watanabe, Y., Ibrahim, M.S., Ellakany, H.F., Kawashita, N., Daidoji, T., Takagi, T., Yasunaga, T., Nakaya, T., and Ikuta, K.
  • Journal Title: J. Gen. Virol. Volume: 93 Issue: 10 Pages: 2215-2226
  • DOI: 10.1099/vir.0.044032-0
  • Peer Reviewed
• [Journal Article] 感染症のメタゲノミック診断 (2011)
  • Author(s): 中村昇太, 中屋隆明, 飯田哲也
  • Journal Title: 化学療法の領域 Volume: 27(No.9) Pages: 114-120
• [Journal Article] Metagenomic analysis of bacterial infections by means of high-throughput DNA sequencing (2011)
  • Author(s): Nakamura S, Nakaya T, Iida T
  • Journal Title: Exp.Biol.Med. Volume: 236(8) Issue: 8 Pages: 968-71
  • DOI: 10.1258/ebm.2011.010378
  • Peer Reviewed
• [Presentation] 新興・再興ウイルス感染症のダイナミクス (2012)
  • Author(s): 中屋隆明
  • Organizer: びわ湖国際医療フォーラム/ 特別講演
  • Place of Presentation: 大津市
  • Year and Date: 2012-07-14
• [Presentation] 次世代シークエンサーが切り拓く感染症のメタゲノム学 (2012)
  • Author(s): 中屋隆明
  • Organizer: 京都産業大学・バイオフォーラム 2011
  • Place of Presentation: 京都市
  • Year and Date: 2012-01-16
• [Presentation] 次世代シークエンサーが切り拓く感染症のメタゲノム (2012)
  • Author(s): 中屋 隆明
  • Organizer: 京都産業大学総合生命科学部第11回バイオフォーラム(招待講演)
  • Place of Presentation: 京都市
• [Presentation] H1N1 2009-HAゲノムのディープシークエンス解析
  • Author(s): 中屋 隆明
  • Organizer: インフルエンザ研究者交流の会シンポジウム
  • Place of Presentation: 福島県耶麻郡
• [Presentation] 新興・再興ウイルス感染症のダイナミクス
  • Author(s): 中屋 隆明
  • Organizer: びわ湖国際医療フォーラム
  • Place of Presentation: 滋賀県大津市
  • Invited
• [Presentation] インフルエンザウイルスの宿主域拡大戦略
  • Author(s): 中屋 隆明
  • Organizer: CREST公開シンポジウム「光が拓く細胞解析の新展開」
  • Place of Presentation: 東京都品川区
• [Presentation] 鳥インフルエンザ
  • Author(s): 中屋 隆明
  • Organizer: 日本神経感染症学会
  • Place of Presentation: 京都府京都市
  • Invited
• [Book] Direct metagenomic detection of viral pathogens in human specimens using an unbiased high-throughput sequencing approach. (2011)
  • Author(s): Nakaya T, Nakamura S, Iida T, Horii Tet al
  • Publisher: Handbook of Molecular Microbial Ecology II, Metagenomics in Different Habitats. Wiley-Blackwell, Hoboken, NJ, USA
• [Book] Handbook of Molecular Microbial Ecology II (2011)
  • Author(s): Nakaya T, Nakamura S, Iida T, Horii T.
  • Total Pages: 609
  • Publisher: Wiley-Blackwell, Hoboken, NJ, USA
• [Remarks]
  • URL: http://www.biken.osaka-u.ac.jp/act/act_nakaya.php
• [Remarks]
  • URL: http://www.f.kpu-m.ac.jp/doc/classes/basicmedicine/221.html
• [Remarks]
  • URL: http://www.f.kpu-m.ac.jp/k/did/