JP6909305B2 - ヌクレアーゼ酵素を産生する微生物の迅速な同定のための方法 - Google Patents
ヌクレアーゼ酵素を産生する微生物の迅速な同定のための方法 Download PDFInfo
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- C12N9/14—Hydrolases (3)
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- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
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Description
‐ 金ナノ粒子とDNA/RNA分子を含む液体に微生物を播種する。一方、この内容では、ヌクレアーゼ酵素を生成する生物は、液体で観察される可能性が高い変色によって検出される。
‐ DNA/RNA分子の一方の末端に蛍光発生分子(フルオロフォア)を結合し、もう一方の末端に蛍光消光(クエンチャー)分子を結合すること、
‐ 前記蛍光発生分子及び前記蛍光消光分子を有する前記DNA/RNA分子を含む溶液を調製し、成長培地へ前記溶液を添加すること、
‐ 前記蛍光発生分子及び前記蛍光消光分子を有する前記DNA/RNA分子を含む前記溶液から成る前記成長培地に、同定する微生物を播種すること、そして、
‐ 光学装置を用いて微生物の前記成長中に前記成長培地中の蛍光シグナルをモニタリングすること。
‐ 正に帯電した(正の)金ナノ粒子と負に帯電した(負の)DNA/RNA分子で構成される溶液を調製すること、
‐ このように調製された溶液に同定される微生物を追加すること、そして、
‐ 溶液に発生する可能性のある変色を監視することである。
(1) Gerceker D, Karasartova D, Elyurek E, Barkar S, Kiyan M, Ozsan TM, Calgin MK, Sahin F. 2009. A new, simple, rapid test for detection of DNase activity of microorganisms: DNase Tube test. J Gen Appl Microbiol. 55 (4): 291-4.
Claims (3)
- 成長中にヌクレアーゼ酵素を産生する微生物をリアルタイムでモニタリングし、迅速に同定する方法であって、以下のステップを含むことを特徴とする。
‐ DNA/RNA分子の一方の末端に蛍光発生分子(フルオロフォア)を結合し、もう一方の末端に蛍光消光(クエンチャー)分子を結合すること、
‐ 前記蛍光発生分子及び前記蛍光消光分子を有する前記DNA/RNA分子を含む溶液を調製し、成長培地へ前記溶液を添加すること、
‐ 前記蛍光発生分子及び前記蛍光消光分子を有する前記DNA/RNA分子を含む前記溶液から成る前記成長培地に、同定する前記微生物を播種すること、そして、
‐ 光学装置を用いて微生物の前記成長中に前記成長培地中の蛍光シグナルをモニタリングすること。 - 成長中にヌクレアーゼ酵素を産生する微生物をリアルタイムでモニタリングし、迅速に同定するために、請求項1による方法を実行するため使用するように構成されたキットであって、以下のものからなることを特徴とする。
・ 適切な生理学的pH値を有し、蛍光発生分子及び蛍光消光分子を有する前記DNA/RNA分子を含む前記溶液から成る前記成長培地、
・ 蛍光研究に適したチューブ及び/或いはプレート、
・ 微生物の前記成長に使用される振盪または非振盪インキュベーター、
・ 前記微生物の播種後、微生物の前記成長中に発生する可能性が高い前記蛍光シグナルを検出及び評価するための光学装置、
・ 前記蛍光シグナルの評価用のソフトウェアを備えた装置。 - 成長中にヌクレアーゼ酵素を産生する微生物をリアルタイムでモニタリングし、迅速に同定するためのキットであって、以下のものからなることを特徴とする。
・ 適切な生理学的pH値を有し、蛍光発生分子及び蛍光消光分子を有するDNA/RNA分子を含む溶液から成る成長培地、
・ 蛍光研究に適したチューブ及び/或いはプレート、
・ 微生物の前記成長に使用される振盪または非振盪インキュベーター、
・ 前記微生物の播種後、微生物の前記成長中に発生する可能性が高い蛍光シグナルを検出及び評価するための光学装置、
・ 前記蛍光シグナルの評価用のソフトウェアを備えた装置。
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TR201703901 | 2017-03-14 | ||
TR2017/03901 | 2017-03-14 | ||
PCT/IB2018/051664 WO2018167666A1 (en) | 2017-03-14 | 2018-03-13 | Method for rapid identification of microorganisms producing nuclease enzymes |
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US (1) | US20200017898A1 (ja) |
EP (1) | EP3596228B1 (ja) |
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WO2021092110A1 (en) * | 2019-11-05 | 2021-05-14 | Nuclease Probe Technologies, Inc. | Microbial detection platform |
EP4209596A1 (en) | 2022-01-07 | 2023-07-12 | Bioanalizes sistemos, UAB | Electrochemical system for the rapid and sensitive detection and quantification of nuclease-like activity exhibiting enzymes |
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CA1118328A (en) * | 1978-05-01 | 1982-02-16 | Christopher Hill | Device for detecting deoxyribonuclease production |
DE3534927A1 (de) | 1985-10-01 | 1987-04-02 | Boehringer Mannheim Gmbh | Phosphate von resorufin-derivaten, verfahren zu deren herstellung sowie deren verwendung zur bestimmung der aktivitaet von phosphatasen |
US6673616B1 (en) | 1992-12-07 | 2004-01-06 | Third Wave Technologies, Inc. | Methods and compositions for characterizing nucleic acids |
DE19843873A1 (de) | 1998-09-25 | 2000-03-30 | Univ Halle Wittenberg | Verfahren zur Bestimmung von Proteaseaktivitäten auf Zelloberflächen |
US20040175737A1 (en) | 2002-12-23 | 2004-09-09 | Wyeth | Assay for RNase H activity |
FR2956868B1 (fr) | 2010-03-01 | 2014-01-10 | Bio Rad Pasteur | Procede rapide de detection d'enzymes et de microorganismes |
CN101871008B (zh) | 2010-05-31 | 2013-07-31 | 浙江工商大学 | 真菌酸性核酸酶检测试剂、制备方法及真菌酸性核酸酶的检测方法 |
CN102321759B (zh) | 2011-08-25 | 2013-05-08 | 南京邮电大学 | 一种检测s1核酸酶及其抑制剂的荧光方法 |
WO2013033436A1 (en) * | 2011-09-01 | 2013-03-07 | University Of Iowa Research Foundation | Oligonucleotide-based probes for detection of bacterial nucleases |
US9632085B2 (en) * | 2012-02-29 | 2017-04-25 | President And Fellows Of Harvard College | Rapid antibiotic susceptibility testing |
WO2014207515A1 (en) | 2013-06-25 | 2014-12-31 | Tubitak (Turkiye Bilimsel Ve Teknolojik Arastirma Kurumu) | Fluorescence method for detecting nuclease activity |
EP3102704B1 (en) * | 2014-02-07 | 2020-02-12 | University of Iowa Research Foundation | Oligonucleotide-based probes and methods for detection of microbes |
ITTO20141040A1 (it) * | 2014-12-15 | 2016-06-15 | Fondazione St Italiano Tecnologia | Procedimento per la rivelazione colorimetrica di contaminazione da nucleasi |
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US20200017898A1 (en) | 2020-01-16 |
JP2020511129A (ja) | 2020-04-16 |
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EP3596228B1 (en) | 2022-09-28 |
WO2018167666A1 (en) | 2018-09-20 |
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