Time-saving method for directly amplifying and capturing a minimal amount of pancreatic tumor-derived mutations from fine-needle aspirates using digital PCR （微量組織検体中の体細胞変異の検出を可能にする迅速デジタルPCR解析法の開発）

Persistent ID (NDL): info:ndljp/pid/11929633

Material type: 博士論文

Author: 林, 明宏

Publisher: -

Publication date: 2020-06

Material Format: Digital

Capacity, size, etc.: -

Name of awarding university/degree: 旭川医科大学,博士（医学）

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Note (General)：: It is challenging to secure a cytopathologic diagnosis using minute amounts of tumor fluids and tissue fragments. Hence, we developed a rapid, accurat...

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Material Type: 博士論文
ISSN: 2045-2322
Title: Time-saving method for directly amplifying and capturing a minimal amount of pancreatic tumor-derived mutations from fine-needle aspirates using digital PCR （微量組織検体中の体細胞変異の検出を可能にする迅速デジタルPCR解析法の開発）
Volume: 10 1
Author/Editor: 林, 明宏
Publication Date: 2020-06
Publication Date (W3CDTF): 2020-06
Periodical title: Scientific reports
Degree grantor/type: 旭川医科大学
Date Granted: 2021-03-25
Date Granted (W3CDTF): 2021-03-25
Dissertation Number: 甲第557号
Degree Type: 博士（医学）
Conferring No. (Dissertation): 甲第557号
Text Language Code: eng
Note (General): It is challenging to secure a cytopathologic diagnosis using minute amounts of tumor fluids and tissue fragments. Hence, we developed a rapid, accurate, low-cost method for detecting tumor cell-derived DNA from limited amounts of specimens and samples with a low tumor cellularity, to detect KRAS mutations in pancreatic ductal carcinomas (PDA) using digital PCR (dPCR). The core invention is based on the suspension of tumor samples in pure water, which causes an osmotic burst; the crude suspension could be directly subjected to emulsion PCR in the platform. We examined the feasibility of this process using needle aspirates from surgically resected pancreatic tumor specimens (n = 12). We successfully amplified and detected mutant KRAS in 11 of 12 tumor samples harboring the mutation; the positive mutation frequency was as low as 0.8%. We used residual specimens from fine-needle aspiration/biopsy and needle flush processes (n = 10) for method validation. In 9 of 10 oncogenic KRAS pancreatic tumor samples, the "water-burst" method resulted in a positive mutation call. We describe a dPCR-based, super-sensitive screening protocol for determining KRAS mutation availability using tiny needle aspirates from PDAs processed using simple steps. This method might enable pathologists to secure a more accurate, minimally invasive diagnosis using minute tissue fragments.
identifier:PMID:32704002
開始ページ : 12332
Persistent ID (NDL): info:ndljp/pid/11929633
https://dl.ndl.go.jp/pid/11929633
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
https://dl.ndl.go.jp/collections/A00014
Acquisition Basis: 博士論文（自動収集）
Date Accepted (W3CDTF): 2021-12-07T08:33:55+09:00
Format (IMT): application/pdf
Access Restrictions: 国立国会図書館内限定公開
Service for the Digitized Contents Transmission Service: 図書館・個人送信対象外
Availability of remote photoduplication service: 可
Periodical Title (URI): https://amcor.asahikawa-med.ac.jp/modules/xoonips/detail.php?id=20210325_K557
Data Provider (Database): 国立国会図書館 : 国立国会図書館デジタルコレクション
https://dl.ndl.go.jp