成果報告書詳細
管理番号20140000000452
タイトル*平成25年度中間年報 「がん超早期診断・治療機器の総合研究開発 超早期高精度診断システムの研究開発:血液中のがん分子・遺伝子診断を実現するための技術・システムの研究開発 血中分子・遺伝子診断のための基礎技術の研究開発 (がんの発症を予測するシステムの開発 乳がん感受性評価システムの研究開発)」
公開日2014/7/26
報告書年度2013 - 2013
委託先名東洋紡株式会社
プロジェクト番号P10003
部署名バイオテクノロジー・医療技術部
和文要約
英文要約Title:Research and development of a new system of risk assessment for breast cancer.(FY2013-FY2014)FY2013 Annual Report

1 Development of CNV measurement reagent
For CNV measurement, the DNA array method is generally used, however, real-time PCR method with Taqman-probe is also often used. Q-probe, which is adopted by the Toyobo’s full automatic genetic analyzer  GENECUBE , is a homogeneous fluorescence assay method similar to Taqman-probe. It is expected to make the reagent at a low cost by adopting the Q-probe and to reduce labor cost by the automatic instrumentation.First, to trace Yamaguchi university experiments results, the three breast cancer susceptibility CNV candidate was measured by real-time PCR method with Taqman-probe, and it was confirmed that CNV can be measured. Then, as an absolute quantitative method, the digital PCR system is also set up at TOYOBO. Based on the above results, we examined the changing to the Q-probe from Taqman-probe. Because Q-probe is based on the G quenching, Q-probe was designed to ensure that there is no G sequences around. Furthermore, dose ratios of PCR primer are optimized for asymmetric PCR. By optimizing the PCR conditions, the amplification curves equivalent to taqman-probe was obtained. Finally, for three breast cancer susceptibility CNV candidates, it was confirmed that CNV values measured by Taqman-probe are in good agreement with those measured by Q-probe and those measured by digital PCR.
2 Development of CNV measurement system
TOYOBO’s GENECUBE is a full automatic genetic analyzer for performing the steps of 1、 DNA extraction from blood samples,2、 setting up for PCR reaction mixture,3、 PCR reaction, and 4、 analysis of PCR amplicon. Furthermore, it can complete analysis in about 1 hour. However, since  GENECUBE  is at the moment a qualitative PCR device, it is necessary for CNV measurement to modify it to be a quantitative PCR device. Therefore, while maintaining the advantages of high speed and full automaticity of GENECUBE, we have planed the remodeling concept to the novel quantitative PCR apparatus.
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