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ID 117206
タイトル別表記
バイサルファイト変換とARMS PCRを用いた膵β細胞傷害の新規定量法
Detecting pancreatic β-cell cfDNA
著者
岡田, 朝美 徳島大学大学院医学研究科(医学専攻)
山田, 美鈴 Tokushima University
Tominaga, Yukari Tokushima University
Jo, Kyoka Tokushima University
Suzuki, Reiko Tokushima University
Akehi, Yuko Tokushima University
髙士, 祐一 Tokushima University
Koga, Daisuke Otsuka Pharmaceutical Co., Ltd.
下北, 英輔 Tokushima University
Mitsui, Yukari Tokushima University
Masuda, Shiho Tokushima University
Ferreri, Kevin City of Hope
Fujitani, Yoshio Gunma University
キーワード
cell free DNA
DNA methylation
Quantitative RT-PCR
amplification-refractry mutation system PCR
Type 1 diabetes
資料タイプ
学位論文
抄録
Aims/Introduction: Several research groups have reported methods for quantifying pancreatic beta cell (β-cell) injury by measuring β-cell-specific CpG unmethylation of the insulin gene in circulation using digital droplet PCR or next-generation sequencing. However, these methods have certain disadvantages, such as the need to consider the background signal owing to the small number of target CpG sites and the need for unique equipment.
Materials and Methods: We established a novel method for detecting four CpG unmethylations of the insulin gene using two-step amplification refractory mutation system PCR. We applied it to type 1 diabetes (T1D) patients with a wide range of disease durations and to healthy adults.
Results: The assay showed high linearity and could detect a single copy of unmethylated insulin DNA in experiments using methylated and unmethylated plasmid DNA. The unmethylated insulin DNA level in the type 1 diabetes group, whose β-cell mass was considerably reduced, was similar to that of healthy adults. An inverse correlation was observed between copy number and disease duration in patients with unmethylated insulin DNA-positive type 1 diabetes.
Conclusions: We developed a novel method for detecting unmethylated insulin DNA in circulation that can be performed using a conventional real-time PCR system. This method would be useful for analyzing dynamic profiles of β-cells in human disease such as type 1 diabetes.
掲載誌名
Journal of Diabetes Investigation
ISSN
20401124
cat書誌ID
AA12488319
出版者
Asian Association for the Study of Diabetes|Wiley
13
7
開始ページ
1140
終了ページ
1148
発行日
2022-04-09
備考
内容要旨・審査要旨・論文本文の公開
本論文は,著者Asami Okadaの学位論文として提出され,学位審査・授与の対象となっている。
権利情報
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License ( https://creativecommons.org/licenses/by-nc/4.0/ ), which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
EDB ID
出版社版DOI
出版社版URL
フルテキストファイル
言語
eng
著者版フラグ
博士論文全文を含む
文科省報告番号
甲第3641号
学位記番号
甲医第1535号
学位授与年月日
2022-06-23
学位名
博士(医学)
学位授与機関
徳島大学
部局
先端酵素学研究所
医学系
生物資源系
病院