新規な遺伝子検査プロトコール開発とアルコール代謝関連遺伝子多型への応用  [in Japanese] Development of Novel Genotyping Protocol and Its Application for Genotyping of Alcohol Metabolism-related Genes  [in Japanese]

Access this Article

Author(s)

    • 今井 美穂 Imai Miho
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室 School of Pharmaceutical Sciences, Mukogawa Women's University
    • 競 和佳 Kisoi Madoka
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室 School of Pharmaceutical Sciences, Mukogawa Women's University
    • 坂口 友唯 Sakaguchi Yui
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室 School of Pharmaceutical Sciences, Mukogawa Women's University
    • 山村 美和子 Yamamura Miwako
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室 School of Pharmaceutical Sciences, Mukogawa Women's University
    • 村田 成範 Murata Shigenori
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室|武庫川女子大学バイオサイエンス研究所 School of Pharmaceutical Sciences, Mukogawa Women's University|Institute of Biosciences, Mukogawa Women's University
    • 市川 厚 Ichikawa Atsushi
    • 武庫川女子大学バイオサイエンス研究所|一般社団法人生命科学教育研究所 Institute of Biosciences, Mukogawa Women's University|Bio Education Laboratory
    • 木下 健司 Kinoshita Kenji
    • 武庫川女子大学薬学部健康生命薬科学科ゲノム機能解析学研究室|武庫川女子大学バイオサイエンス研究所|一般社団法人生命科学教育研究所 School of Pharmaceutical Sciences, Mukogawa Women's University|Institute of Biosciences, Mukogawa Women's University|Bio Education Laboratory

Abstract

<p>A new single nucleotide polymorphisms (SNP) genotyping method has been developed and validated using biological specimens directly as templates for TaqMan PCR without general DNA extraction and purification procedure from dried saliva samples attached on water-soluble papers. This new method can set up at ease and complete PCR analysis including data interpretation in under two hours with additional advantages of application for large-scale clinical research, diagnostics, and epidemiological studies at low cost. Specifically, SNP genotyping of alcohol metabolism-related genes <i>ADH1B</i> (rs1229984) and <i>ALDH2</i> (rs671) were demonstrated by TaqMan PCR assay using dried saliva samples in the present investigation. In this protocol, by simplifying experimental operations and improving efficiency, omitting and simplifying the time and laborious DNA purification process, it is possible to shorten the experiment time and reduce the risk of human error such as contamination. Furthermore it became possible with great cost reduction. We succeeded in dramatically improving the judgment rate and accuracy of SNP genotyping by the master mix reagent for commercial available real-time TaqMan PCR. Moreover, it becomes possible to stably introduce template DNA into the reaction system, and it will be possible to apply it to copy number variation (CNV) by TaqMan probe method. The SNP analysis process using this optimized water-soluble paper will be applied to gene polymorphism analysis of drug metabolizing enzyme gene CYP, <i>etc.</i>, to help efforts to realize personalized medicine.</p>

Journal

  • YAKUGAKU ZASSHI

    YAKUGAKU ZASSHI 139(8), 1111-1119, 2019

    The Pharmaceutical Society of Japan

Codes

  • NII Article ID (NAID)
    130007686242
  • NII NACSIS-CAT ID (NCID)
    AN00284903
  • Text Lang
    JPN
  • ISSN
    0031-6903
  • NDL Article ID
    029922779
  • NDL Call No.
    Z19-411
  • Data Source
    NDL  J-STAGE 
Page Top