JP2020511129A - 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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- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases RNAses, DNAses
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- G01N2333/90—Enzymes; Proenzymes
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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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Abstract
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/蛍光分子(フルオロフォア)を、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、
‐ そのようなDNA/RNA分子を含む溶液を調製し、前述の物を培地に入れること、
‐ 同定する微生物を培地に追加すること、そして、
‐ 光学装置によって培地中の蛍光シグナルのモニタリング。
‐ 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 (6)
- 本発明は、ヌクレアーゼ酵素を産生する微生物を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、
‐ そのようなDNA/RNA分子を含む溶液を調製し、前の物を培地に入れること、
‐ 同定する微生物を培地に播種すること、そして、
‐ 光学装置を用いて培地中の蛍光シグナルのモニタリング。 - 本発明は、ヌクレアーゼ酵素を産生する微生物を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。
‐ 正に帯電した金ナノ粒子と負に帯電したDNA/RNA分子で構成される溶液を調製すること、
‐ 同定する微生物を溶液に追加すること、そして、
‐ 溶液の変色のモニタリング。 - 請求項1に記載のヌクレアーゼ酵素を産生する微生物の迅速な同定方法であって、適切な生理学的pH値を有する培地が使用されるキットを含むことを特徴とする。
- 請求項3に記載のヌクレアーゼ酵素を産生する微生物の迅速な同定方法であって、キット、蛍光研究に適したチューブ及び/或いはプレート、微生物の増殖に使用される振盪または非振盪インキュベーター、微生物の播種後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、そしてシグナルの評価用のソフトウェアが使用される装置を含むことを特徴とする。
- 請求項2に記載のヌクレアーゼ酵素を産生する微生物の迅速な同定方法であって、正に荷電した金ナノ粒子及び負に荷電したDNA/RNA分子からなり、適切なpH値を有する溶液を含むキットを含むことを特徴とする。
- 請求項5に記載のヌクレアーゼ酵素を産生する微生物の迅速な同定方法であって、キット、変色に適したチューブ及び/或いはプレート、微生物の播種後に生じる変色に関連する吸収シグナルを検出及び評価する光学装置。また、信号の評価に使用されるソフトウェアを使用する装置を含むことを特徴とする。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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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 |
Publications (2)
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JP2020511129A true JP2020511129A (ja) | 2020-04-16 |
JP6909305B2 JP6909305B2 (ja) | 2021-07-28 |
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JP2019547683A Active JP6909305B2 (ja) | 2017-03-14 | 2018-03-13 | 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 |
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US (1) | US20200017898A1 (ja) |
EP (1) | EP3596228B1 (ja) |
JP (1) | JP6909305B2 (ja) |
WO (1) | WO2018167666A1 (ja) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
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US20220380828A1 (en) * | 2019-11-05 | 2022-12-01 | 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 |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
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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 |
EP2751567B1 (en) * | 2011-09-01 | 2019-05-08 | University of Iowa Research Foundation | Oligonucleotide-based probes for detection of bacterial nucleases |
WO2013130875A1 (en) * | 2012-02-29 | 2013-09-06 | 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 |
WO2015120406A1 (en) * | 2014-02-07 | 2015-08-13 | 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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- 2018-03-13 EP EP18720360.9A patent/EP3596228B1/en active Active
- 2018-03-13 JP JP2019547683A patent/JP6909305B2/ja active Active
- 2018-03-13 WO PCT/IB2018/051664 patent/WO2018167666A1/en active Search and Examination
- 2018-03-13 US US16/493,274 patent/US20200017898A1/en not_active Abandoned
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Publication number | Publication date |
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WO2018167666A1 (en) | 2018-09-20 |
EP3596228A1 (en) | 2020-01-22 |
JP6909305B2 (ja) | 2021-07-28 |
EP3596228B1 (en) | 2022-09-28 |
US20200017898A1 (en) | 2020-01-16 |
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