| Project/Area Number |
20300183
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Medical systems
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
LEZHAVA Alexander The Institute of Physical and Chemical Research, オミックス機能研究ユニット, ユニットリーダー (40443048)
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| Co-Investigator(Kenkyū-buntansha) |
OKAMOTO Akimitsu 独立行政法人理化学研究所, 岡本独立主幹研究ユニット, ユニットリーダー(独立主幹研究員) (60314233)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥19,630,000 (Direct Cost: ¥15,100,000、Indirect Cost: ¥4,530,000)
Fiscal Year 2010: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2009: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
Fiscal Year 2008: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
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| Keywords | SmartAmp法 / Exciton Primer / SNPタイピング / Cancer Mutation Detection / Multiplex reaction / End Product Visualization / Warfarin / EGFR / Smart Amplification Process / SNP genotyping / Mutation detection / Two-color reaction / Visualization of amplified product / Cancer / Virus Detection / エキシトンプライマー / 増幅検出の可視化 / マルチプレックス / 遺伝子増幅 / 等温増幅 / 新規蛍光物質 / 目視判定 / multiplex |
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| Research Abstract |
A two-color readout of the Smart Amplification (SmartAmp、SMAP) genotyping system using so-called Exciton Primers have been developed. New method is highly specific and sensitive, permitting a one-step, single-tube mutation detection reaction which delivers a result in 45 minutes directly from blood. Exciton Primers, which are functioning as "fluorescence emission with sequence-specific," after hybridization to complementary sequences, can provide the signals resulting target sequence detection for real-time monitoring of amplification reactions. Applied to the isothermal SmartAmp2 mutation detection process, Exciton Primers show high signal strength with low background leading to a superior specificity and sensitivity compared to SYBR Green I. The genotyping assay can use only one labeled Exciton Primer for endpoint detection, or simultaneously by real-time monitoring detect wild-type and mutant alleles in a one-tube reaction using two Exciton Primers having different dyes. The technique seems very promising for decentralized point-of-care diagnostics since the approach does not involve complicated instrumentation and sample purification. We demonstrated efficiency of the new technology on the example of rapid detection of SNPs and mutations
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