Mixture analysis and age estimation based on mitochondrial DNA polymophisms and methylation pattern

• Project/Area Number: 17K09259
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Legal medicine
• Research Institution: Asahikawa Medical College
• Principal Investigator: Masaru Asari 旭川医科大学, 医学部, 准教授 (40360979)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 個人識別 / 混合試料 / ミトコンドリアDNA / メチル化 / 法医学 / 混合DNA / 年齢推定 / ミトコンドリアDNA多型

Outline of Final Research Achievements

We developed a new method for discrimination of mitochondrial DNA haplotypes based on inhibition of amplification using designed probes including single nucleotide polymorphisms or reference sequences. Our method could effectively discriminate each haplotype from mitochondrial DNA mixtures. Although we did not find the useful methylation targets for age estimation of Japanese individuals, we confirmed that single bese extention method would be useful to detect lower methylation rates.

Academic Significance and Societal Importance of the Research Achievements

本研究結果は、きわめて微量な混合試料からのミトコンドリアDNA型解析法として法医実務へ利用可能である。特に検体数が多い場合においても、効率よく被疑者および被害者など含まれる個人のミトコンドリアDNA型が検出可能であると考える。

Research Products

• [Journal Article] Discrimination of haplotype in mitochondrial DNA mixtures using LNA-mediated PCR clamping. (2019)
  • Author(s): Asari M, Isozaki S, Hoshina C, Okuda K, Tanaka H, Horioka K, Shiono H, Shimizu K.
  • Journal Title: Forensic Sci Int Genet. Volume: 41 Pages: 58-63
  • DOI: 10.1016/j.fsigen.2019.03.018
  • Peer Reviewed
• [Presentation] PCRクランピング法を用いた混合試料からのミトコンドリアDNA型解析 (2019)
  • Author(s): 浅利 優，磯崎翔太郎，保科千里，奥田勝博，田中宏樹，北村麻奈，島津雅子，堀岡希衣，塩野 寛，清水惠子
  • Organizer: 第103次日本法医学会学術全国集会
• [Presentation] 浅利 優，奥田勝博，保科千里，田中宏樹，堀岡希衣，塩野 寛，清水惠子 (2019)
  • Author(s): 配列特異的な増幅阻害に基づいた混合試料からのミトコンドリアDNA型解析
  • Organizer: 日本DNA多型学会第28回学術集会