Development of the novel high precision low labor load gene-diagnosis system towards realization of tailor-made medical care

• Project/Area Number: 23550092
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: University of Fukui
• Principal Investigator: TAKAHASHI Toru 福井大学, 工学(系)研究科(研究院), 講師 (30361166)
• Co-Investigator(Kenkyū-buntansha): IKI Nobuhiko 東北大学, 大学院・環境科学研究科, 准教授 (50282108)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 一塩基多型 / キャピラリー電気泳動 / 一塩基多型検出 / ジェノタイピング / SNP検出 / 抽出ゲノム / PCR / SNP / 一塩基変異一本鎖DNA / 塩基 / プロトン解離 / 分離 / 検出 / 判定法

Research Abstract

A multiple detection method for single nucleotide polymorphism (SNP) based on the CE separation technique for single base substituted sequence isomers of single stranded DNA was proposed. In the CE separation method where an acidic aqueous urea solution was employed as an electrophoretic buffer solution, the separation of single base substituted sequence isomers of single stranded DNA was achieved according to the difference of the proton dissociation properties of each DNA bases at acidic condition. A clear criterion for determining of existence or nonexistence of SNP by the peak pattern of the electropherogram allows high reliability of the proposed method. The proposed method can be applied to all substitution patterns with identical condition. The proposed method was successfully applied to the SNP detection and genotyping for extracted human gene fragment sample including ALDH2 gene region.

Research Products

• [Journal Article] Dynamic Coating Capillary Electrophoresis for Separation of Humic Acid Using Mixture Solution of Non-ionic Polymers both as Coating Agent and Separation Medium (2013)
  • Author(s): T. Takahashi, J. Kawana, Y. Tamura, H. Hoshino
  • Journal Title: Anal. Sci Volume: 29 Pages: 1099-1102
• [Journal Article] Dissociation Kinetic Analysis of Ce(III) Complex with Quin2 by Microchip Capillary Electrophoretic Reactor (2013)
  • Author(s): K. Ohtsuka, N. Iki, H. Hoshino, T. Takahashi
  • Journal Title: Anal. Sci Volume: 29 Pages: 553-557
  • NAID: 10031170985
• [Journal Article] Dynamic Coating Capillary Electrophoresis for Separation of Humic Acid Using Mixture Solution of Non-ionic Polymers both as Coating Agent and Separation Medium (2013)
  • Author(s): T. Takahashi, J. Kawana, Y. Tamura, H. Hoshino
  • Journal Title: Analytical Sciences Volume: 29 Issue: 11 Pages: 1099-1102
  • DOI: 10.2116/analsci.29.1099
  • NAID: 130004442170
  • ISSN: 0910-6340, 1348-2246
  • Peer Reviewed
• [Journal Article] Dissociation Kinetic Analysis of Ce(III) complex with Quin2 by Microchip Capillary Electrophoretic Reactor (2013)
  • Author(s): Kei-ichirou OHTSUKA, Nobuhiko IKI, Hitoshi HOSHINO, and Toru TAKAHASHI
  • Journal Title: Anal. Sci Volume: 29 Pages: 553-557
  • NAID: 10031170985
  • Peer Reviewed
• [Journal Article] Highly Efficient Near-Infrared-Emitting Lanthanide(III) Complexes Formed by Heterogeneous Self-Assembly of AgI, LnIII, and Thiacalix[4]arene-p-tetrasulfonate in Aqueous Solution (LnIII = NdIII, YbIII) (2012)
  • Author(s): Iki, Nobuhiko; Hiro-oka, Shouichi; Tanaka, Teppei; Kabuto, Chizuko; Hoshino, Hitoshi
  • Journal Title: Inorganic Chemistry Volume: 51 Pages: 1648-1656
  • Peer Reviewed
• [Presentation] ハイブリダイズによらない一塩基多型検出およびジェノタイピング : 一本鎖DNAのキャピラリー電気泳動分離による一塩基変異の直接識別 (2013)
  • Author(s): 高橋 透, 深川 真有, 櫻井 隆郎, 星野 仁
  • Organizer: 第34回キャピラリー電気泳動シンポジウム
  • Place of Presentation: 日本女子大学目白キャンパス
• [Presentation] 一本鎖DNAのキャピラリー電気泳動分離に基づく一塩基多型検出法とジェノタイピングへの応用 (2013)
  • Author(s): 高橋 透, 深川 真有, 櫻井 隆郎, 星野 仁
  • Organizer: 日本分析化学会第62回年会
  • Place of Presentation: 近畿大学東大阪キャンパス
• [Presentation] ハイブリダイズによらない一塩基多型検出およびジェノタイピング：一本鎖DNAのキャピラリー電気泳動分離による一塩基変異の直接識別 (2013)
  • Author(s): 高橋 透，深川 真有，櫻井 隆郎，星野 仁
  • Organizer: 第34回キャピラリー電気泳動シンポジウム
  • Place of Presentation: 日本女子大学目白キャンパス
• [Presentation] A novel multi-target SNP detection method based on the capillary electrophoresis separation technique for single base substituted single-stranded DNA (2011)
  • Author(s): Toru Takahashi, Takao Sakurai
  • Organizer: International Congress on Analytical Chemistry 2011 (ICAS2011)
  • Place of Presentation: 京都国際会議場
• [Presentation] 2.キャピラリー電気泳動分離に基づく天然高分子のキャラクタリゼーション法の開発 (2011)
  • Author(s): 高橋 透
  • Organizer: 第31 回キャピラリー電気泳動シンポジウム(招待講演)
  • Place of Presentation: 慶応大学鶴岡キャンパス
• [Presentation] A novel multi-target SNP detection method based on the capillary electrophoresis separation technique for single base substituted single-stranded DNA (2011)
  • Author(s): T. Takahashi, T. Sakurai
  • Organizer: International Congress on Analytical Sciences 2011
  • Place of Presentation: 京都国際会議場
• [Book] in Fundamental Concepts, Practical Applications, and Limitations of Capillary Electrophoresis and Microchip Capillary Electrophoresis, C. D. Garcia, K. Chumbimuni Torres, and E. Carrilho (Eds.), Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor : Dissociation Kinetic Analysis Method for "Complexes" Using Capillary Electrophoretic Separation Process (2013)
  • Author(s): T. Takahashi, N. Iki
  • Publisher: John Wiley & Sons, Inc
• [Book] 5. “Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor: Dissociation Kinetic Analysis Method for “Complexes” Using Capillary Electrophoretic Separation Process” , in Fundamental Concepts, Practical Applications, and Limitations of Capillary Electrophoresis and Microchip Capillary Electrophoresis (2013)
  • Author(s): Toru Takahashi, Nobuhiko Iki
  • Total Pages: 394
  • Publisher: John Wiley & Sons, Inc.
• [Book] Capillary Electrophoretic Reactor and Microchip Capillary Electrophoretic Reactor: Dissociation Kinetic Analysis Method for "Complexes" Using Capillary Electrophoretic Separation Process, in C. D. Garcia, K. Chumbimuni Torres, and E. Carrilho (Eds.), Fundamental Concepts, Practical Applications, and Limitations of Capillary Electrophoresis and Microchip Capillary Electrophoresis (2012)
  • Author(s): T. Takahashi, N. Iki
  • Publisher: John Wiley & Sons