Development of Digital Isothermal Nucleic Acid Amplification for Rapid Quantification
| Project/Area Number |
16K15334
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Laboratory medicine
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| Research Institution | University of Yamanashi (2018)
Institute of Physical and Chemical Research (2016-2017) |
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Principal Investigator |
Tanaka Yuji 山梨大学, 大学院総合研究部, 特任准教授 (40625513)
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| Co-Investigator(Kenkyū-buntansha) |
寺尾 泰久 順天堂大学, 医学部, 先任准教授 (00348997)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Keywords | 等温核酸増幅 / デジタル / 逆転写 / ゲノム変異 / 融解曲線解析 / 再生医療 / 網膜再生 / 等温増幅 / 遺伝子変異 / RPE / 核酸マーカー / iPS細胞 / 網膜色素上皮細胞 / デジタル等温増幅法 / 子宮体癌 / 迅速診断 / デジタル核酸定量 / 迅速核酸定量 / 診断技術 / バイオマーカー |
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| Outline of Final Research Achievements |
In this study, we examined two new detection methods: digital nucleic acid quantification method with reverse transcription and digital nucleic acid quantification method of identifying and quantifying nucleic acid amplified by melting curve analysis. The isothermal amplification method can amplify a nucleic acid only by maintaining it at around 70 ° C, but there is a problem that it can not be determined unless there is a 10-fold concentration difference, but digitization of the isothermal amplification kit with reverse transcription It has been found that it becomes possible to detect even a concentration difference of about 2 times. In addition, in order to detect genomic mutations used for quality control of cells utilized for regenerative medicine of retina, newly designed primers for isothermal amplification, digital nucleic acid amplification, and biotyped and mutated nucleic acid by melting curve analysis. It turned out that it can be identified and quantified.
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| Academic Significance and Societal Importance of the Research Achievements |
ゲノムをあらかじめ調べた上でその患者に適した薬剤を処方する「個別化医療」の実用化が始まり、より簡易的に、より正確に、より迅速に、より低価格で、より多様な核酸の差異を識別するための技術開発の重要性が高まっている。本研究でRNAの迅速簡易定量とゲノム変異の融解曲線解析による定量という２つの新たな検出方法の可能性を見出した。既存の核酸検出の改良や、新たな個別化医療や再生医療の創出に貢献する可能性がある。
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Report
(4 results)
Research Products
(14 results)
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[Journal Article] Promoter-level transcriptome in primary lesions of endometrial cancer identified biomarkers associated with lymph node metastasis.2017
Author(s)
Yoshida E, Terao Y, Hayashi N, Mogushi K, Arakawa A, Tanaka Y, Ito Y, Ohmiya H, Hayashizaki Y, Takeda S, Itoh M, Kawaji H.
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Journal Title
Sci Rep
Volume: 7(1):14160
Issue: 1
Pages: 1-15
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] RIKEN Symposium: CAGE as a tool for cancer research and biomarker discovery2016
Author(s)
Kaczkowski Bogumil, Tanaka Yuji, Kawaji Hideya, Andersson Robin, Albin Sandelin, Itoh Masayoshi, Lassmann Timo, the FANTOM5 consortium, Hayashizaki Yoshihide, Carninci Piero, Forrest Alistair Raymond Russel.
Organizer
RIKEN Symposium : Cancer research by CAGE method
Place of Presentation
Riken Yokohama Koryuto-hall, Yokohama, Japan
Year and Date
2016-09-13
Related Report
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[Presentation] Novel approach to predict lymph node metastatic state of uterine endometrial cancer, toward reduction of lymphedema by detection of irrelevant lymphadenectomy2016
Author(s)
Emiko Yoshida, Yasuhisa Terao, Noriko Hayashi, Yosuke Ito, Kazunari Fujino, Takafumi Ujihira, Soshi Kusunoki, Miki Kimura, Hiroshi Kaneda, Tsuyoshi Ota, Daiki Ogishima, Kiyoko Kato, Yuji Tanaka, Masayoshi Ito, Hideya Kawaji, Atsuo Itakura, Satoru Takeda
Organizer
第58回 日本婦人科腫瘍学会学術講演会
Place of Presentation
米子
Year and Date
2016-07-08
Related Report
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[Presentation] CLST Educational Program: Cap Analysis of Gene Expression (CAGE) and cancer biology2016
Author(s)
Kaczkowski Bogumil, Tanaka Yuji, Kawaji Hideya, Andersson Robin, Albin Sandelin, Itoh Masayoshi, Lassmann Timo, the FANTOM5 consortium, Hayashizaki Yoshihide, Carninci Piero, Forrest Alistair Raymond Russel
Organizer
CLST Educational Program
Place of Presentation
RIKEN Kobe MI R&D Center , KOBE, Japan
Year and Date
2016-07-05
Related Report
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[Presentation] Transcriptome analysis of recurrently deregulated genes across multiple cancers identifies new pan-cancer biomarkers., American Association for Cancer Research2016
Author(s)
Kaczkowski Bogumil, Tanaka Yuji, Kawaji Hideya, Andersson Robin, Albin Sandelin, Itoh Masayoshi, Lassmann Timo, the FANTOM5 consortium, Hayashizaki Yoshihide, Carninci Piero, Forrest Alistair Raymond Russel
Organizer
Annual Meeting 2016,
Place of Presentation
Ernest N. Morial Convention Center, New Orleans, United States
Year and Date
2016-04-16
Related Report
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