Novel method utilizing bisulfite conversion with dual amplification-refractory mutation system polymerase chain reaction to detect circulating pancreatic β-cell cfDNA

• 資料種別: 博士論文
• タイトル: Novel method utilizing bisulfite conversion with dual amplification-refractory mutation system polymerase chain reaction to detect circulating pancreatic β-cell cfDNA
• 著者・編者: Okada, Asami; Yamada-Yamashita, Misuzu; Tominaga, Yukari; Jo, Kyoka; Mori, Hiroyasu; Suzuki, Reiko; Ishizu, Masashi; Tamaki, Motoyuki; Akehi, Yuko; Takashi, Yuichi; Koga, Daisuke; Shimokita, Eisuke; Tanihara, Fuminori; Kurahashi, Kiyoe; Yoshida, Sumiko; Mitsui, Yukari; Masuda, Shiho; Endo, Itsuro; Aihara, Ken-ichi; Kagami, Shoji; Abe, Masahiro; Ferreri, Kevin; Fujitani, Yoshio; Matsuhisa, Munehide; Kuroda, Akio
• 著者標目: Okada, Asami; Yamada-Yamashita, Misuzu; Tominaga, Yukari; Jo, Kyoka; Mori, Hiroyasu; Suzuki, Reiko; Ishizu, Masashi; Tamaki, Motoyuki; Akehi, Yuko; Takashi, Yuichi; Koga, Daisuke; Shimokita, Eisuke; Tanihara, Fuminori; Kurahashi, Kiyoe; Yoshida, Sumiko; Mitsui, Yukari; Masuda, Shiho; Endo, Itsuro; Aihara, Ken-ichi; Kagami, Shoji; Abe, Masahiro; Ferreri, Kevin; Fujitani, Yoshio; Matsuhisa, Munehide; Kuroda, Akio
• 出版事項: Asian Association for the Study of Diabetes|Wiley
• 出版年月日等: 2022-04-09
• 出版年（W3CDTF）: 2022-04-09
• 並列タイトル等: 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
• タイトル（掲載誌）: Journal of Diabetes Investigation
• 巻号年月日等（掲載誌）: 13 7
• 掲載巻: 13
• 掲載号: 7
• 掲載ページ: 1140-1148
• ISSN（掲載誌）: ISSN : 2040-1124
• 授与機関名: 徳島大学
• 授与年月日: 2022-06-23
• 報告番号: 甲第3641号
• 学位: 博士（医学）
• 本文の言語コード: eng
• 件名標目: cell free DNA; DNA methylation; Quantitative RT-PCR; amplification-refractry mutation system PCR; Type 1 diabetes
• 対象利用者: 一般
• 一般注記: 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.
• 記録形式（IMT）: application/pdf
• オンライン閲覧公開範囲: インターネット公開
• 著作権情報: 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.
• 関連情報（DOI）: 10.1111/jdi.13806
• 連携機関・データベース: 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）; https://irdb.nii.ac.jp
• 提供元機関・データベース: 徳島大学 : 徳島大学機関リポジトリ

目次

http://repo.lib.tokushima-u.ac.jp/files/public/11/117206/20220713155323224695/k3641_abstract.pdf fulltext

http://repo.lib.tokushima-u.ac.jp/files/public/11/117206/20220713155323540432/k3641_review.pdf fulltext

http://repo.lib.tokushima-u.ac.jp/files/public/11/117206/20220713155323828514/k3641_fulltext.pdf fulltext