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METHOD AND KIT FOR DETECTING PATHOGENIC MICROORGANISM

外国特許コード F180009402
整理番号 (AF19P018)
掲載日 2018年4月20日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2017JP031689
国際公開番号 WO 2018043733
国際出願日 平成29年9月4日(2017.9.4)
国際公開日 平成30年3月8日(2018.3.8)
優先権データ
  • 特願2016-172515 (2016.9.5) JP
  • 特願2017-099579 (2017.5.19) JP
発明の名称 (英語) METHOD AND KIT FOR DETECTING PATHOGENIC MICROORGANISM
発明の概要(英語) Provided is a method for detecting pathogenic microorganisms in a biological sample, which is a technique that can be used to perform high-sensitivity detection of pathogenic microorganisms, such as influenza virus, etc., the method including: an introducing step for introducing a hydrophilic solvent that contains the biological sample and substances that serve as substrates for reactions involving enzymes that are present on surfaces of the pathogenic microorganisms or in the interiors thereof into a space between a lower-layer section in which a plurality of accommodating sections that can accommodate the pathogenic microorganisms are formed and an upper-layer section that faces a surface in which the accommodating sections are formed in the lower-layer section; an encapsulating step for introducing a hydrophobic solvent into the space and forming, in the accommodating sections, droplets of the hydrophilic solvent that are coated by the hydrophobic solvent enveloping the pathogenic microorganisms and the substances; and a detecting step for optically detecting reaction products that are generated as a result of the reactions between the enzymes and the substances in the droplets, wherein the hydrophilic solvent has a pH value that is greater than the acid dissociation constant (pKa) of the reaction products.
従来技術、競合技術の概要(英語) BACKGROUND ART
In recent years, simple immunoaffinity chromatography using a test kit is influenza virus has been developed (see Patent Document 1). The method of using immunoaffinity chromatography, from a few minutes to several tens of minutes the influenza virus can be detected, such as diagnosis and treatment of infections are utilized.
In addition, the prior art, the influenza virus and neuraminidase is an enzyme chromogenic substrate and optically based on the reaction of the detection of an influenza virus has been known a technique (Patent Document 2, reference 3). Colored substrates include, for example, 4-- α-D - methylumbellerferyl (4-Methylumbelliferyl-N-acetyl- α-D-neuraminic acid neuramatic acid: 4MU-NANA, see Patent Document 2) or, 4-or 4,7 - alkoxy -N- - N - dialkoxy-acetylneuraminic acid (see Patent Document 3) and derivatives thereof-acetylneuraminic acid and the like is used. For example, as a method using a chromogenic substrate is 4MU-NANA, neuraminidase by the decomposition of the fluorescent substance by 4MU-NANA 4-methylumbelliferone is generated. The generated 4-based on the amount of enzymatic activity of the neuraminidase methylumbelliferone value can be calculated, based on the enzymatic activity of the influenza virus further particle number can be quantified.
Associated with the present invention, the non-patent document 1, and covered with the oil droplets, the droplets from the outside of the femtoliter order accessible using the array of droplets, a method of an enzyme molecule has been described.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • NOJI Hiroyuki
  • TABATA Kazuhito
国際特許分類(IPC)
参考情報 (研究プロジェクト等) CREST Creation of Nanosystems with Novel Functions through Process Integration AREA
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