Development of a PCR/LDR/Flow-Through Hybridization Assay Using a Capillary Tube, Probe DNA-Immobilized Magnetic Beads and Chemiluminescence Detection

• HOMMATSU Manami

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

• OKAHASHI Hisamitsu

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

• OHTA Keisuke

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

• TAMAI Yusuke

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

• TSUKAGOSHI Kazuhiko

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

• HASHIMOTO Masahiko

  Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

抄録

A polymerase chain reaction (PCR)/ligase detection reaction (LDR)/flow-through hybridization assay using chemiluminescence (CL) detection was developed for analyzing point mutations in gene fragments with high diagnostic value for colorectal cancers. A flow-through hybridization format using a capillary tube, in which probe DNA-immobilized magnetic beads were packed, provided accelerated hybridization kinetics of target DNA (i.e. LDR product) to the probe DNA. Simple fluid manipulations enabled both allele-specific hybridization and the removal of non-specifically bound DNA in the wash step. Furthermore, the use of CL detection greatly simplified the detection scheme, since CL does not require a light source for excitation of the fluorescent dye tags on the LDR products. Preliminary results demonstrated that this analytical system could detect both homozygous and heterozygous mutations, without the expensive instrumentation and cumbersome procedures required by conventional DNA microarray-based methods.

収録刊行物

• Analytical Sciences

  Analytical Sciences 29 (7), 689-695, 2013

  社団法人 日本分析化学会

キーワード

• Point mutation; polymerase chain reaction (PCR)
• ligase detection reaction (LDR)
• hybridization
• capillary
• magnetic bead
• chemiluminescence detection

詳細情報

• CRID: 1390001204259141632
• NII論文ID: 10031185229
• NII書誌ID: AA10500785
• DOI: 10.2116/analsci.29.689
• COI: 1:STN:280:DC%2BC3sjpvVaksQ%3D%3D
• ISSN: 13482246
• ISSN: 09106340
• NDL書誌ID: 024671746
• PubMed: 23842410
• Web Site: http://id.ndl.go.jp/bib/024671746
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I024671746
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.29.689.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.29.689/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/29/7/29_689/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2014028719
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可