| Project/Area Number |
17K08988
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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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| Research Field |
Laboratory medicine
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| Research Institution | University of Miyazaki |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
岡山 昭彦 宮崎大学, 医学部, 教授 (70204047)
越本 知大 宮崎大学, フロンティア科学総合研究センター, 教授 (70295210)
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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 |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 細菌検査 / レーザーマイクロダイセクション / Laser Microdissection / 細菌同定 / 16S rRNA / 感染症 / 臨床検査医学 |
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| Outline of Final Research Achievements |
To detect causative microorganisms is important to select accuracy treatments in daily clinical practice of infectious diseases. Gram staining is a common technique to estimate causative microorganisms. However, sometimes, it is difficult to distinguish bacterial contamination of clinical samples in this method. In the present study, we tried to establish a novel method to detect "true" causative microorganisms using laser capture microdissection (LCM). Bacteria-phagocytized neutrophils were targeted and were collected by LCM. After nucleic acid extraction from these neutrophils, the bacteria-specific 16S ribosomal RNA was analyzed. In peritonitis model mice, a causative microorganism was detected by this method. Therefore, LCM method may be a useful tool to detect causative microorganism which was phagocytized by neutrophils. In future study, it is necessary to evaluate the sensitivity and specificity of this method in clinical samples.
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| Academic Significance and Societal Importance of the Research Achievements |
喀痰や尿などの臨床検体における起炎菌の同定は，抗菌薬の選択を適切に行うために重要であるが，日常診療で取り扱う臨床検体はしばしば起炎菌以外の細菌に汚染されており，判断が難しい場合がある．好中球に貪食された細菌は起炎菌と考えられるため，本研究ではこの様な好中球をレーザーキャプチャーマイクロダイセクション（LCM）法で効率的に回収し，起炎菌を同定する方法の開発を進めた．グラム染色とLCM法を組み合わせ，細菌を貪食した好中球の回収と細菌特異的な16SリボゾームRNA遺伝子の同定が可能となった．腹膜炎モデル動物においても本方法により起炎菌の同定が可能であり，今後，臨床検体での応用が期待される．
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