| Project/Area Number |
17K19849
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Society medicine, Nursing, and related fields
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| Research Institution | Yokkaichi Nursing and Medical Care University (2022)
Gihu University of Medical Science (2017) |
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Principal Investigator |
Hoshino Mari 四日市看護医療大学, 看護医療学部, 教授 (10313511)
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| Co-Investigator(Kenkyū-buntansha) |
高崎 昭彦 四日市看護医療大学, 看護医療学部, 教授 (40247664)
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| Project Period (FY) |
2017-06-30 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2017: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | Diagnosis / Escherichia coli / 細菌 |
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| Outline of Final Research Achievements |
In this study we aimed to discover new target proteins to detect all pathotypes of enteric pathogenic Escherichia coli using mass spectrometry, and to develop simple, rapid and high sensitive assays including EIA and multiplex PCR. First, we developed a one-step multiplex PCR-based assay for simultaneous detection and classification of virulence factors to identify five diarrheagenic E. coli pathotypes.
Furthermore, we could find out more than 7 target protein candidates, and developed ELISA to detect the first candidate. However, we detected no difference of the amount of the first candidate protein product between pathogenic E. coli and non-pathogenic E. coli.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究で開発検討を行った下痢原性大腸菌の検査法は、遺伝子レベルでは、行政での検査で下痢原性大腸菌の分類指標にしている病原因子遺伝子全てを検出でき、用いた遺伝子群はこれまでのマルチプレックスPCR法の論文で報告をされていない。また、蛋白質レベルでは、これまで全く対象とされていなかった、病原因子以外の新規蛋白質を分類指標として標的とすることに加え、最終目的としてイムノクロマトまで開発できた場合、保健所・衛生研究所・病院の検査室に検体をまわすことなく、高価な試薬、遺伝子解析の技術習得・熟練等を必要とせずに誰でも、その場で迅速・簡便に全種の下痢原性大腸菌の検出・分類ができるようになる利点を持つ。
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