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Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection
http://hdl.handle.net/10458/0002000355
http://hdl.handle.net/10458/0002000355cf5848e4-1546-4f77-b584-df7c4f8ba7d2
| 名前 / ファイル | ライセンス | アクション |
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著者原稿 (11 MB)
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| Item type | 学術雑誌論文 / Journal Article(1) | |||||||
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| 公開日 | 2023-11-28 | |||||||
| タイトル | ||||||||
| タイトル | Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection | |||||||
| 言語 | en | |||||||
| 言語 | ||||||||
| 言語 | eng | |||||||
| キーワード | ||||||||
| 言語 | en | |||||||
| 主題Scheme | Other | |||||||
| キーワード | Gram stain | |||||||
| キーワード | ||||||||
| 言語 | en | |||||||
| 主題Scheme | Other | |||||||
| キーワード | Laser capture microdissection | |||||||
| キーワード | ||||||||
| 言語 | en | |||||||
| 主題Scheme | Other | |||||||
| キーワード | Polymerase chain reaction | |||||||
| 資源タイプ | ||||||||
| 資源タイプ | journal article | |||||||
| 著者 |
山田, 明輝
× 山田, 明輝× 梅木, 一美× 佐伯, 裕二× 橋倉, 悠輝× Nomura, Hajime
× Yamamoto, Ikuo× 梅北, 邦彦× 高城, 一郎× 越本, 知大× 岡山, 昭彦 |
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| 抄録 | ||||||||
| 内容記述タイプ | Abstract | |||||||
| 内容記述 | Although isolation and identification of bacteria in a clinical specimen constitute essential steps for the diagnosis of bacterial infection, positive results of the bacterial culture are not always attained, despite observing the bacteria by Gram staining. As bacteria phagocytosed by the leukocytes are considered as the causative agents of infectious diseases, this study aims to introduce a new approach for the collection of only bacteria phagocytosed by the neutrophils in an animal model using laser capture microdissection (LCM) followed by the DNA identification using polymerase chain reaction (PCR). We inoculated representative bacteria (Escherichia coli and Staphylococcus aureus) into the abdominal cavities of specific pathogen-free C57BL/6 J mice. After 6 h inoculation, we collected the fluid samples from the peritoneal cavities of mice and demonstrated peritonitis by the increase of neutrophils. Then, we smeared the neutrophils on the membrane slides and collected single-cell phagocytosing bacteria by LCM. The supernatant of the cell lysate was supplied for the PCR reaction to amplify the 16S rRNA gene, and we validated the DNA sequences specific for the inoculated bacteria. In addition, PCR using specific primers for E. coli and S. aureus identified each species of bacteria. Hence, this study suggests that the combination of LCM and PCR could be a novel approach to determine bacteria in infectious diseases. Nevertheless, further investigation is warranted to test various additional bacterial taxa to demonstrate the general applicability of this method to clinical samples. | |||||||
| 言語 | en | |||||||
| 内容記述 | ||||||||
| 内容記述タイプ | Other | |||||||
| 内容記述 | Citation: Yamada A, Umeki K, Saeki Y, Hashikura Y, Nomura H, Yamamoto I, Umekita K, Takajo I, Koshimoto C, Okayama A. Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection. J Microbiol Methods. 2018 Dec;155:42-48. doi: 10.1016/j.mimet.2018.11.005. Epub 2018 Nov 10. PMID: 30423364. |
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| 言語 | en | |||||||
| 書誌情報 |
en : Journal of Microbiological Methods 巻 155, p. 42-48, 発行日 2018 |
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| 出版者 | ||||||||
| 出版者 | Elsevier | |||||||
| 言語 | en | |||||||
| ISSN | ||||||||
| 収録物識別子タイプ | ISSN | |||||||
| 収録物識別子 | 01677012 | |||||||
| DOI | ||||||||
| 関連タイプ | isVersionOf | |||||||
| 識別子タイプ | DOI | |||||||
| 関連識別子 | https://doi.org/10.1016/j.mimet.2018.11.005 | |||||||
| 著者版フラグ | ||||||||
| 出版タイプ | AM | |||||||
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山田, 明輝, 梅木, 一美, 佐伯, 裕二, 橋倉, 悠輝, Nomura, Hajime, Yamamoto, Ikuo, 梅北, 邦彦, 高城, 一郎, 越本, 知大, 岡山, 昭彦, 2018, Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection: Elsevier, 42–48 p.
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