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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.
特許請求の範囲(英語) [claim1]
1. Subject infected with a pathogenic microorganism is suspected of having infected with or separated from the object in a biological sample wherein the method to detect a pathogenic microorganism, wherein said pathogenic microorganism is capable of receiving a plurality of storage sections, having an upper surface of the hydrophobic are spaced from each other by the side wall is formed between the lower portion, the lower layer part in the accommodating portion is formed in the surface facing the space between the upper layer portion and, wherein the biological sample, wherein said pathogenic microorganism is present on or in which a reaction of the material as the substrate, including introducing a procedure of a hydrophilic solvent, a hydrophobic solvent is introduced into the space, wherein the stored in the storage unit, wherein the hydrophobic solvent and coated with a substance wherein said pathogenic microorganism, droplets of a solvent to form a hydrophilic encapsulating procedure, wherein the enzyme and the substance in the liquid droplets in to produce a reaction product generated by the reaction of optically detecting the detection procedure, comprises, the content of the hydrophilic solvent, wherein the acid dissociation constant (pKa) of the reaction product has a value greater than the pH, method.
[claim2]
2. The pathogenic organism is a virus, and in which the enzyme neuraminidases in, that said substance is 4-- α-D - (4-Methylumbelliferyl-N-acetyl- methylumbellerferyl neuramatic acid α-D-neuraminic acid) and, said reaction product through 4-is methylumbelliferone, method according to claim 1.
[claim3]
3. Subject infected with a pathogenic microorganism is suspected of having infected with or separated from the object to detect a pathogenic microorganism in said biological sample kit for, pathogenic microorganism that is capable of housing a plurality of storage sections on the upper surface of the side wall having a hydrophobic formed separated from each other by a low layer part and, wherein the lower portion in the accommodating portion is formed to be spaced apart from the surface and the upper layer part on the opposite side of the array comprises, in the surface or inside of a pathogenic microorganism wherein there is a substrate for the reaction and the material as the enzyme, the enzyme and the material of the reaction product generated by the reaction of an acid dissociation constant (pKa) of a hydrophilic solvent having a pH greater than the value, and the hydrophobic solvent, the kit comprising a.
[claim4]
4. The hydrophobic solvent in a hydrophilic solvent to the contact interface, and the enzyme, the enzyme and a substrate for the reaction by reacting a substance, a method for detecting a reaction product, wherein the content of the hydrophilic solvent, wherein the acid dissociation constant (pKa) of the reaction product has a value greater than the pH, method.
[claim5]
5. Wherein the content of the hydrophilic solvent comprises pathogenic microorganisms, in which the enzyme in the surface or inside of a pathogenic microorganism is present in an enzyme cleavage of the substrate, wherein the chromogenic substance and, in which the enzyme Shaukasten by cleavage of the substrate to produce a reaction product generated by the optical detection, the method according to claim 4.
[claim6]
6. The pathogenic organism is a virus, in which the enzyme neuraminidases and, in that the substrate Shaukasten 4-methylumbellerferyl - α-D - neuraminic acid (4-Methylumbelliferyl-N-acetyl- α-D-neuraminic acid) and, said reaction product 4-in methylumbelliferone, method according to claim 5.
[claim7]
7. Wherein the content of the hydrophilic solvent, pathogenic microorganism subject infected subject suspected of being infected or that has been separated from a biological sample, of claims 5 or 6.
  • 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)
指定国 (WO201843733)
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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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