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ペプチド核酸peptide nucleic acid (PNA)とloop-mediated isothermal amplification assay(LAMP)法を組み合わせた、Mycoplasma pneumoniaeの23S rRNA変異の革新的検出法
https://saitama-med.repo.nii.ac.jp/records/636
https://saitama-med.repo.nii.ac.jp/records/6369f0309a7-3d06-4080-ba53-4b1005f2e6cb
名前 / ファイル | ライセンス | アクション |
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論文内容の要旨 (152.3 kB)
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論文審査の結果の要旨 (81.1 kB)
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Item type | 学位論文 / Thesis or Dissertation(1) | |||||
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公開日 | 2018-05-30 | |||||
タイトル | ||||||
タイトル | ペプチド核酸peptide nucleic acid (PNA)とloop-mediated isothermal amplification assay(LAMP)法を組み合わせた、Mycoplasma pneumoniaeの23S rRNA変異の革新的検出法 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | A novel detection procedure for mutations in the 23S rRNA gene of Mycoplasma pneumoniae with peptide nucleic acid-mediated loop-mediated isothermal amplification assay. | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Colorimetry | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | DNA, Bacterial | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Genes, Bacterial | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Genes, rRNA | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Genotype | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Humans | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Limit of Detection | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Mycoplasma pneumoniae | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Nasopharynx | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Nucleic Acid Amplification Techniques | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Peptide Nucleic Acids | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Pneumonia, Mycoplasma | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Point Mutation | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Point-of-Care Testing | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Polymerase Chain Reaction | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | RNA, Ribosomal, 23S | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Sensitivity and Specificity | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Temperature | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_46ec | |||||
資源タイプ | thesis | |||||
著者 |
酒井, 純
× 酒井, 純× Sakai, Jun |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Rapid and easy detection of a single nucleotide point mutation of bacterial genes, which is directly linked to drug susceptibility, is essential for the proper use of antimicrobial agents. Here, we established a detection method using a peptide nucleic acid mediated loop-mediated amplification (LAMP) assay for macrolide (ML)-susceptible Mycoplasma pneumoniae. This assay specifically detected the absence of missense mutations encoding the central loop of domain V in the gene encoding 23S rRNA, which can reduce the affinity for MLs and subsequently generate ML-resistant strains of M. pneumoniae. Reactions were performed at 62°C for 60min and targeted gene amplifications were detected by real-time turbidity with a turbidimeter and naked-eye inspection of a color change. The assay had an equivalent detection limit of 100.0fg of DNA with the turbidimeter and showed specificity against 54 types of pathogens, whereas amplification was completely blocked, even at 100.0pg of DNA per reaction, in the presence of point mutations at 2063A and 2064A. The expected LAMP products were confirmed through identical melting curves in real-time LAMP procedures. This method would be a simple and rapid protocol for single nucleotide polymorphism genotyping as point-of-care testing technology without amplification of the sequences carrying the point mutations 2063A and 2064A in ML-resistant M. pneumoniae strains. | |||||
学位名 | ||||||
学位名 | 博士(医学) | |||||
学位授与機関 | ||||||
学位授与機関名 | 埼玉医科大学 | |||||
学位授与年度 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 平成29年度 | |||||
学位授与年月日 | ||||||
学位授与年月日 | 2018-03-23 | |||||
学位授与番号 | ||||||
学位授与番号 | 32409甲第1384号 | |||||
掲載誌名 | ||||||
関連名称 | Journal of Microbiological Methods | |||||
掲載誌情報 | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://www.sciencedirect.com/science/article/pii/S0167701217302178?via%3Dihub | |||||
関連名称 | 出版社サイト |