| Project/Area Number |
17K18112
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Physical pharmacy
Laboratory medicine
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| Research Institution | Showa University |
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Principal Investigator |
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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| Keywords | miRNA / NASBA / ピロリン酸 / PPDK / イソルミノール化学発光 / POCT / 分析化学 / 臨床検査システム |
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| Outline of Final Research Achievements |
A compact, simple, quick, and high sensitive method is presently required in order to perform simple genetic analysis in a clinic or bedside (Point-of-care test; POCT). Nucleic Acid Sequence-Based Amplification (NASBA) is a specifically amplify method of a single-stranded RNA fragment. The detection by chemiluminescence reaction (CL) is not required a light source. We developed a novel analytical system of miRNA for POCT by combination with NASBA and CL.
In order to detect the NASBA product with high sensitivity, we developed a high sensitive determination method for PPi which amount is correlated with the amount of the NASBA product. We succeeded in the determination of PPi by the optimization of various conditions. Consequently, miR-196a was detected by combining NASBA and CL, and discriminated between negative and positive template. This combined miRNA detection method could be useful for compact POCT system in the future clinical field.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では、簡便なmiRNAのPoint of care testing (POCT)システムを開発した。本法は、定温増幅法であるNASBA法によりターゲットmiRNAを増幅し、同時に生成するピロリン酸をイソルミノール化学発光法により検出する新たな方法である。使用する機器は、手のひらサイズにすることが可能であり、癌や感染症などの簡易検査法として利用することが出来る。さらに単一のmiRNAのみではなく様々なmiRNAに応用可能である。この分析技術を用いて、将来の薬局における薬剤師による受診勧奨の補助、病院や検査所の簡易検査室における定期健診や診断の補助にも利用されることが望まれる。
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