Development of 19F-NMR-Based Multiplex Focused Gene Diagnostic Systems

• Project/Area Number: 24510165
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Nanomaterials/Nanobioscience
• Research Institution: University of Tsukuba
• Principal Investigator: OISHI Motoi 筑波大学, 数理物質系, 講師 (90419242)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
• Keywords: 19Ｆ－ＮＭＲ / ＤＮＡ / 磁性粒子 / 金ナノ粒子 / 19F-NMR / DNA / センサー / 酵素

Outline of Final Research Achievements

An ultrasensitive colorimetric assay for both DNA and RNA has been developed utilizing enzyme-free click chemical ligation chain reaction (CCLCR) on dispersed gold nanoparticles and magnetic beads. The CCLCR assay provides ultrasensitive detection (50 zM = 10-21 M: several copies) of target DNA that is comparable to PCR-based approaches. Note that target RNA could also be detected with similar sensitivity without the need for reverse transcription to the corresponding cDNA. Thus, this method might be a useful tool for development of 19F-NMR–based multiplex focused gene diagnostic systems.

Research Products

• [Journal Article] Enzyme-Free and Isothermal Detection of microRNA Based on Click-Chemical Ligation-Assisted Hybridization Coupled with Hybridization Chain Reaction Signal Amplification (2015)
  • Author(s): Ｍｏｔｏｉ Ｏｉｓｈｉ
  • Journal Title: Analytical and Bioanalytical Chemistry Volume: in press Issue: 14 Pages: 4165-4172
  • DOI: 10.1007/s00216-015-8629-y
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Ultrasensitive Detection of DNA and RNA Based on Enzyme-Free Click Chemical Ligation Chain Reaction on Dispersed Gold Nanoparticles (2014)
  • Author(s): Daiki Kato and Motoi Oishi
  • Journal Title: ACS Nano Volume: 8 Issue: 10 Pages: 9988-9997
  • DOI: 10.1021/nn503150w
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Enzyme-Free Fluorescent-Amplified Aptasensors Based on Target-Responsive DNA Strand Displacement via Toehold-Mediated Click Chemical Ligation (2014)
  • Author(s): Motoi Oishi, Shingo Nakao, and Daiki Kato
  • Journal Title: Chemical Communications Volume: 50 Pages: 991-993
  • Peer Reviewed
• [Journal Article] Rational Design of a Fluorescence-Amplified Aptasensor Based on Enzyme-Assisted Target Recycling Strategy (2013)
  • Author(s): Shingo Nakao and Motoi Oishi
  • Journal Title: Chemistry Letters Volume: 42 Issue: 10 Pages: 1122-1124
  • DOI: 10.1246/cl.130439
  • NAID: 10031201282
  • Peer Reviewed
• [Presentation] DNAクリックケミカルライゲーションを用いたmiRNAの検出 (2014)
  • Author(s): 大石 基, 高島明里
  • Organizer: 日本分析化学会第63年会
  • Place of Presentation: 広島大学
  • Year and Date: 2014-09-17 – 2014-09-19
• [Presentation] 金ナノ粒子表面での酵素フリー型クリックケミカルライゲーション連鎖反応を基盤としたDNAおよびRNAの超高感度検出システムの構築 (2014)
  • Author(s): 加藤大喜, 大石 基
  • Organizer: 第74回日本分析化学討論会
  • Place of Presentation: 日本大学、郡山
  • Year and Date: 2014-05-24 – 2014-05-25
• [Presentation] 金ナノ粒子表面での酵素フリー型クリックケミカルライゲーション連鎖反応を基盤としたDNAおよびRNAの超高感度検出システムの構築 (2014)
  • Author(s): 大石 基
  • Organizer: 第74回分析化学討論会
  • Place of Presentation: 福島県郡山市 日本大学工学部
• [Presentation] DNAおよびRNAの超高感度検出を目指した金ナノ粒子表面での酵素フリー (2014)
  • Author(s): 大石 基
  • Organizer: ナノ学会 第12回
  • Place of Presentation: 京都府宇治市 京都大学宇治キャンパス
• [Presentation] 酵素および酵素フリーでのシグナル増強を目指した蛍光アプタマーセンサーの構築 (2013)
  • Author(s): 大石 基
  • Organizer: 日本分析化学会第62年会
  • Place of Presentation: 大阪府東大阪市 近畿大学東大阪キャンパス
• [Presentation] 構造スイッチングDNAアプタマー用いた酵素増強型蛍光バイオセンサーの創製 (2013)
  • Author(s): 大石 基
  • Organizer: 第73回分析化学討論会
  • Place of Presentation: 北海道函館市、北海道大学函館キャンパス
• [Presentation] スマートナノデバイスによるガンのピンポイント診断・治療システムの開発
  • Author(s): 大石 基
  • Organizer: 第４回つくばグリーンイノベーションフォーラム「バイオサイエンス」
  • Place of Presentation: 茨城県つくば市、筑波大学