| Project/Area Number |
18K11680
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 64010:Environmental load and risk assessment-related
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| Research Institution | University of Miyazaki |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
真砂 佳史 国際連合大学サステイナビリティ高等研究所, サステイナビリティ高等研究, その他 (50507895)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 水系感染症 / 病原細菌 / 病原遺伝子 / 一斉検出・定量 / マルチプレックスPCR / NGSシーケンシング / コピー数 / リード数 / プライマー設計 / 一斉検出 / 次世代シーケンサー / 泡沫濃縮法 / 都市河川 / 菌叢解析 / 病原性細菌属 / 感染リスク / 河川水 / 病原細菌濃縮 / 細菌同時検出 |
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| Outline of Final Research Achievements |
Waterborne diseases due to pathogenic bacteria in the water environment are a serious problem all over the world. Therefore, it is necessary to develop a method that can accurately and simultaneously obtain information on pathogenic bacteria in the water environment. In this study, the development of a detection method that combines the simultaneous amplification of target genes using the Multiplex PCR method and the simultaneous analysis of base sequences by the NGS analysis were investigated for seven types of E. coli pathogenic genes. As a result of the Multiplex PCR method, it was possible to simultaneously detect 5 out of 7 target genes. The relationship between the copy number of the digital PCR method established as a quantitative analysis and the read number of the NGS analysis, five types of E. coli pathogen-ic genes showed a positive correlation. In addition, it was suggested that the copy number can be estimated from the number of reads.
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| Academic Significance and Societal Importance of the Research Achievements |
病原性細菌の検出技術が発展しているにも関わらず，水環境において，それら病原性細菌の一斉検出法は確立されておらず，情報・知見が乏しいのが現状である．本研究では，Multiplex PCR法における多数の標的遺伝子を一斉に増幅させるプロセスとNGS法による塩基配列の一斉解析のプロセスを組み合わせて，水環境における病原性細菌の一斉検出法の開発した．この開発した方法によって，これまで得られていなかった水環境における病原遺伝子の存在実態と分布が追跡できる．
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