成果報告書詳細
管理番号20120000001196
タイトル*平成23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・遺伝子診断自動化システムの研究開発(血中がん遺伝子診断の検体処理自動化システム)」
公開日2012/12/19
報告書年度2011-2011
委託先名コニカミノルタテクノロジーセンター株式会社、プレシジョン・システム・サイエンス株式会社、株式会社ファインラバー研究所
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
和文要約和文要約等以下本編抜粋:
1.共同研究の内容及び成果等
(1) 血液検体評価機能の開発(担当:コニカミノルタテクノロジーセンター株式会社)
1.目的
末梢血中の循環がん細胞(CTC)を高感度に検出する技術を開発する。抗体と細胞表面抗原との反応を利用した免疫学的手法を用いた方法などが開発されているが、検出感度が十分でない。特に、抗体と反応が弱い細胞種が検出できないことや、検体性状が測定結果に影響しやすいことなどが問題となっている。そこで本研究開発では、検体に極微量に存在するCTCを細胞表面抗原に依存せず、できる限りロスや損傷なく、含有率や性状を高感度に同定するシステムを開発することを目的とする。
英文要約Title: Development of Automatic Testing System for Genetic Diagnosis using Peripheral Blood (FY2010-FY2012) FY2011 Annual Report
1) Development of an evaluation system for blood samples (Konica Minolta Technology Center, Inc.)
An accurate inspection system detecting less than 100 CTCs in 10 ml peripheral blood has been developed. In each process, the following results are obtained. (1) Preparation of blood samples: Density-gradient centrifugation was found to be a useful way to reduce the numbers of RBCs to the same level of WBCs even a few days after collecting the blood. (2) Alignment and immunostaining of cells on a cell chip: A slide glass size chip on which lots of micro-well chambers were fabricated was introduced to retain a great number of cells. A controlled flow of the blood sample allowed the chip hold approximately two million cells in monolayer. Further, we confirmed that fluorescent immunostaining to the retained cells worked appropriately. (3) Optical instruments for detecting rare cells: To identify CTCs, we optimized optical and signal processing conditions, and then set to design the prototype for performance confirmation of the whole system.
2) Development of a sample preparation system for genetic diagnostics (by Precision System Science Co., Ltd.) It is necessary to standardize the sample preparation procedure and to establish its evaluation method for its practical use in genetic diagnosis. The development of an automated system is indispensable for reliability and accuracy. We aimed here to develop an automated system with a RNA extraction function along with its evaluation function. The challenge to the minimization of the extension board was continued. Succeeded in downsizing of approximately 28%, was able to be achieved to reduce the size of the extraction instrument. And we fabricated a prototype of the conceptual design and then integrated a process of nucleic acid analysis by micro-TAS followed by nucleic acid extraction purification step. The prototype instrument should be installed an optimized rapid RNA extraction protocol with pre-filled reagents and also liquid handling protocol for RT-PCR.
3) Development of sample evaluation system for genetic diagnostics (by Fine Rubber R&D Co., Ltd.)
We established a lamination technology for decrease laminating error via optimized laminating condition to manufacture micro-TAS device made of silicone rubber sheets. We developed non-adhesive preparation chemical agent to make non-adhesive areas for flow channels or valve mechanism of micro-TAS device. And to resolved screen printing method for method of non-adhesive preparation to silicone rubber sheets with non-adhesive preparation chemical agent and adopted a printing machine. We selected the cultured cell which made cirrhosis the origin to evaluate the efficiency of the micro-TAS device. That performance evaluation is compared with the gene expression profile data acquired with a micro-array. And we fabricated the conceptual design and prototype of an integrated process and the concatenation of nucleic acid analysis by micro-TAS and nucleic acid extraction/purification step.
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