Simple and rapid detection of Rothia mucilaginosa by loop-mediated isothermal amplification method

Persistent ID (NDL): info:ndljp/pid/12071567

Material type: 博士論文

Author: 中井, 啓介

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Publication date: 2018-03-09

Material Format: Digital

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Name of awarding university/degree: 大阪歯科大学,博士（歯学）

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Note (General)：: Rothia mucilaginosa is an emerging opportunistic pathogen in immunocompromised hosts. The aim of this study was to develop a rapid and simple examinat...

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Material Type: 博士論文
Title: Simple and rapid detection of Rothia mucilaginosa by loop-mediated isothermal amplification method
Author/Editor: 中井, 啓介
Author Heading: 中井, 啓介
Publication Date: 2018-03-09
Publication Date (W3CDTF): 2018-03-09
Alternative Title: LAMP 法を応用した Rothia mucilaginosa の迅速・簡易検出法の確立
Degree grantor/type: 大阪歯科大学
Date Granted: 2018-03-09
Date Granted (W3CDTF): 2018-03-09
Dissertation Number: 甲第828号
Degree Type: 博士（歯学）
Conferring No. (Dissertation): 甲第828号
Text Language Code: eng
Subject Heading: Rothia mucilaginosa
LAMP
naked eye inspection
bacteremia
Target Audience: 一般
Note (General): Rothia mucilaginosa is an emerging opportunistic pathogen in immunocompromised hosts. The aim of this study was to develop a rapid and simple examination method for this pathogen.The loop-mediated isothermal amplification (LAMP) method was applied. Two primer sets (FIP, BIP, F3, B3 and two loop primers) were designed including species-specific sequences of the 16S ribosomal RNA gene. The LAMP reactions were performed under isothermal conditions at 64°C for 20 or 50 minutes. The target gene was specifically amplified by LAMP with one primer set. Unexpectedly, the other primer set showed nonspecific amplification for other Rothia species. The amplicons were visualized by agarose gel electrophoresis with detection limits of 10 and 100 cells/tube for the 50-minute and 20-minute LAMP methods, respectively. By addition of SYBR-Green I, naked eye inspection was possible with the same detection limit. To further simplify the method, intact cells were subjected to the LAMP assay, and the amplicons were detected with a limit of 10 cells in the 50-minute reaction.Using LAMP, we developed a simple and rapid detection assay for R. mucilaginosa. This assay may be useful for microbiological diagnosis.
2017年度
Persistent ID (NDL): info:ndljp/pid/12071567
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Collection: 障害者向け資料
Collection (Materials For Handicapped People:1): テキストデータ
Collection (particular): 国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文
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Acquisition Basis: 博士論文（自動収集）
Date Accepted (W3CDTF): 2022-02-06T04:33:19+09:00
Date Created (W3CDTF): 2018-05-17
Format (IMT): application/pdf
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Periodical Title (URI): http://id.nii.ac.jp/1392/00000171/
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