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CELL PROLIFERATION DETECTION METHOD USING FLUORESCENCE IN WATER-IN-OIL EMULSION CULTURING コモンズ

外国特許コード F190009840
整理番号 (2017002067)
掲載日 2019年7月25日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2018JP037257
国際公開番号 WO 2019073902
国際出願日 平成30年10月4日(2018.10.4)
国際公開日 平成31年4月18日(2019.4.18)
優先権データ
  • 特願2017-197980 (2017.10.11) JP
発明の名称 (英語) CELL PROLIFERATION DETECTION METHOD USING FLUORESCENCE IN WATER-IN-OIL EMULSION CULTURING コモンズ
発明の概要(英語) The present invention addresses the problem of providing a method which enables the noninvasive detection and isolation of droplets in which microorganisms have proliferated. The present invention relates to a detection method for detecting the proliferation of microorganisms within droplets, the method including (a) a step for producing droplets which include microorganisms, (b) a step for culturing the microorganisms within the droplets, and (c) a step for detecting proliferation of the microorganisms within the droplets, in which (c1) a fluorescent modified nucleic acid probe, which acts on a substance secreted or discharged from the microorganisms and emits fluorescence, is used, and fluorescence emitted by the fluorescent modified nucleic acid probe is measured or (c2) the autofluorescence of the microorganisms is measured, wherein the droplets in which fluorescence was detected include intact microorganisms.
従来技術、競合技術の概要(英語) BACKGROUND ART
Into the environment and various organisms, we have produced by the microorganism or microorganisms these materials have been used. However, in the environment 99% is not yet successful culture, many of the resources of the organism remains untouched. These can be the unknown microorganism is cultured, a physiological characteristic of the microorganisms in the environment as well as a clear, medical, the development in the industry is expected to be applied. In recent years, separation of the microorganisms, as one of the culture, the method using the emulsion W/O being of interest (non-patent document 1 and Non-Patent Document 2). In this manner, the water in the oil phase (microbial culture medium used) is dispersed, each of the droplets (droplet) is used as one of the culture field. By using the micro-channels, several hundred thousand to several tens of minutes-several hundred million units of the droplet can be manufactured and high throughput can be expected. In addition, the number of microorganisms present in the aqueous phase by adjusting the number of droplet, the droplets enclosed in one cell can be produced, from a single cell culture is enabled (non-patent document 3). When culturing a microorganism utilizing this approach, the number of cells per one droplet 1 according to the Poisson distribution. 1 Enters the one cell to one of the droplet increases the probability of the substrate increases, there is no probability of occurrence of microbes of the droplet increases. At this time, it can be seen that the growth of microorganisms selectively droplet as long as the sorting of each microorganism analysis, can be used to scale up and the like. The detection of microorganisms in the droplet present in the cell reacts with the substrate fluorescence have been used, these cells are disrupted at the same time is required because, while living microorganisms cannot be taken out (non-patent document 4, Non-Patent Document 5).
  • 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
  • NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
  • 発明者(英語)
  • OOTA Yuri
  • SAITO Kanako
  • TAKAGI Taeko
  • TSUNEDA Satoshi
  • MIYAMOTO Tatsuki
  • MORITA Masamune
  • MATSUKURA Satoko
  • NODA Naohiro
国際特許分類(IPC)
指定国 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 JP 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) Biomedical Research Institue,Bioanalytical Research Group

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