JP5475278B2 - 細胞内酵素放出前の細胞外酵素不活性化を用いたサンプル中の細胞識別 - Google Patents
細胞内酵素放出前の細胞外酵素不活性化を用いたサンプル中の細胞識別 Download PDFInfo
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Description
(a)前記サンプルが:(i)測定可能な活性を有する酵素を含む細胞外培地を含んでなり;且つ(ii)前記測定可能な活性を有する酵素を細胞内に有する標的細胞を含む疑いがあり;
(b)前記方法が:(i)前記細胞外培地内の前記測定可能な活性は不活性化するが、前記標的細胞中の前記測定可能な活性は不活性化しない試薬で、前記液体サンプルを処理する工程;(ii)前記標的細胞を溶解して前記細胞内酵素を放出させる工程;及び(iii)前記測定可能な活性を測定する工程を含んでなる方法を提供する。
(a)前記サンプルが:(i)測定可能な活性を有する酵素を含む細胞外培地を含んでなり;且つ(ii)前記測定可能な活性を有する細胞内酵素を含む標的細胞を含む疑いがあり;
(b)前記方法が:(i)前記細胞外培地中の前記測定可能な活性を不活性化するが、前記標的細胞中の前記測定可能な活性を不活性化しない試薬で、前記液体サンプルを処理する工程;(ii)工程(i)後に残存する前記標的細胞の培養物を確立する工程;並びに、(iii)前記培養物から1又は2以上のサンプルを取得する工程を含んでなる。
(a)前記サンプルが:(i)測定可能な活性を有する酵素を含む細胞外培地を含んでなり;且つ(ii)前記測定可能な活性を有する細胞内酵素を含む標的細胞を含む疑いがあり;
(b)前記方法が:
(i)微生物の増殖を許容する条件下で前記サンプルを温置する工程;
(ii)第1の時点において、前記サンプルから第1の下位サンプルを取得する工程;
(iii)細胞外培地中の前記測定可能な活性を不活性化するが、前記標的細胞中の前記測定可能な活性を不活性化しない試薬で、前記第1の下位サンプルを処理する工程;
(iv)前記標的細胞を溶解して前記細胞内酵素を放出させる工程;
(v)前記測定可能な活性を測定する工程;
(vi)第2の時点において、前記サンプルから第2の下位サンプルを取得する工程;
(vii)その細胞外培地中の前記測定可能な活性を不活性化するが、前記標的細胞中の前記測定可能な活性を不活性化しない試薬で、前記第2の下位サンプルを処理する工程;
(viii)前記標的細胞を溶解し、前記細胞内酵素を放出させる工程;及び、
(ix)前記測定可能な活性を測定する工程を含んでなり、前記第1の時点と前記第2の時点との間で測定可能な活性に増加が見られる場合には、これらの時点間で前記標的細胞が増殖したことが示される、方法が提供される。
抗微生物剤を所定の濃度でサンプルに加える工程;
前記抗微生物剤の存在下、前記サンプルを所定の期間(例えば、抗微生物剤の不在下で>2対数の増殖が見られる期間)に亘って温置する工程;及び
前記期間の終わりにサンプル中の微生物数を調べる工程を有する。
複数の異なる抗微生物剤を所定の濃度で、複数の異なる下位サンプルに加える工程;
前記下位サンプルを前記抗微生物剤の存在下、所定の期間に亘って温置する工程;及び
前記期間の終わりに、前記下位サンプル中の微生物数を調べる工程を有する。
抗微生物剤を複数の所定の濃度で、複数の異なる下位サンプルに加える工程;
前記下位サンプルを前記抗微生物剤の存在下、所定の期間に亘って温置する工程;及び
前記期間の終わりに前記下位サンプル中の微生物数を調べる工程を有する。
抗微生物剤を所定の濃度で下位サンプルに加える工程;
前記下位サンプルを前記抗微生物の存在下、所定の期間に亘って温置する工程;及び
前記期間内の複数の時点で前記下位サンプル中の微生物数を調べる工程を有する。
(a)前記サンプルが:(i)細胞外培地中の前記測定可能な活性を不活性化するが、前記標的細胞中の前記測定可能な活性は不活性化しない試薬で処理された液体サンプルを含んでなり;且つ(ii)前記測定可能な活性を有する細胞内酵素を含む標的細胞を含む疑いがあるものであって;
(b)前記方法が:(i)前記標的細胞を溶解して前記細胞内酵素を放出させる工程;及び(ii)前記測定可能な活性を含んでなる方法を提供する。
(a)前記サンプルが:(i)測定可能な活性を有する酵素を含む細胞外培地を含んでなり;且つ(ii)前記測定可能な活性を有する細胞内酵素を含む標的細胞を含んでいる疑いがあり;
(b)前記方法が:(i)前記細胞外培地中の前記測定可能な活性を不活性化するが、前記標的細胞中の前記測定可能な活性を不活性化しない試薬で、液体サンプルを処理する工程を含んでなる方法を提供する。
a.10mgの16,000U/mgトリプシンを16mlのブロスに加え、10,000U/ml(10×)のストックを得る。
b.25mgの4U/mgパパインを1mlのブロスに加え、100U/ml(10×)のストックを得る。
c.10mgの7.5U/mgプロテイナーゼKを3mlのブロスに加え、25U/ml(10×)のストックを得る。
d.10.5μlの239U/mlキモトリプシンを239.5μlのブロスに加え、10U/mlのストックを得る。
[1]http://www. altcorp. com/AffinityLaboratory/introgcf.htm
[2]WO94/17202
[3]WO96/02665
[4]Squirrell et al. (1994) Adenylate kinase as a cell marker in bioluminescent assays. In: Campbell AK, Kricka LJ, Stanley PE (editors). Bioluminescence and Chemiluminescence: Fundamentals and Applied Aspects. Chichester: John Wiley and Sons, pages 486-9
[5]Sanders (1994) A rapid bioluminescent technique for the detection and identification of Listeria monocytogenes in the presence of Listeria innocua. Pages 454-7 of same volume as for ref. 4
[6]WO00/70082
[7]Blasco et al. (1998) Journal of Applied Microbiology
[8]WO00/46357
[9]WO99/23489
[10]Bean et al. (1992) InternationalJournal of Cell Cloning 10:190-5
[11]Konstantinova (1972) Dokl Akad Nauk SSSR 203:1204-6
[12]Perrier et al. (1998) J Biol Chem 273:19097-101
[13]Fraser et al. (1997) Biochem J 323:711-8
[14]Balows (1974) Current techniques for antibiotic susceptibility testing. ISBN: 0398028869
[15]Smaill (2000) Can J Gastroenterol 14:871-875
[16]Baquero (1990) Eur J Clin Microbiol Infect Dis 9:492-495
[17]Baselski (1996) Clin Lab Med. 16:49-60
[18]Kassimi et al. (2002) J Virol Methods 101:197-206
[19]Zhang & Weintraub (1998) J Clin Microbiol 36:3545-3548
[20]Dowd et al. (1998) Appl Environ Microbiol 64:333-336
[21]Li et al. (1996) J Clin Microbiol 34:1903-1907
[22]Shetab et al. (1998) J Clin Microbiol 36:1729-1732
[23]Homes et al. (1991) J Clin Microbiol 29:2375-2379
Claims (3)
- 液体サンプル中における標的細胞の存否を検出する方法であって:
(a)前記サンプルが:(i)測定可能な活性を有する酵素を含む細胞外培地を含んでなり;且つ(ii)前記測定可能な活性を有する酵素を細胞内に有する標的細胞を含む疑いがあり;
(b)前記方法が:(i)前記細胞外培地内の前記測定可能な活性は不活性化するが、前記標的細胞中の前記測定可能な活性は不活性化しない試薬で、前記液体サンプルを処理する工程;(ii)前記標的細胞を溶解して前記細胞内酵素を放出させる工程;及び(iii)前記測定可能な活性を測定する工程を含んでなり、
ここで、前記測定可能な活性を有する酵素がアデニル酸キナーゼを含んでなり、前記細胞外培地内の前記測定可能な活性を不活性化する試薬が酸又はプロテアーゼを含んでなる、方法。 - 液体サンプルが血液サンプルであり、前記酵素がアデニル酸キナーゼである請求項1に記載の方法に使用されるキットであって、
前記工程(b)(i)においてアデニル酸キナーゼを不活性化するための試薬であるプロテアーゼ;
前記工程(b)(ii)において血液細胞を溶解するための試薬であるサポニンと;
ADP;
ルシフェリン;及び
ルシフェラーゼ
を含んでなるキット。 - 液体サンプルが血液サンプルであり、前記酵素がアデニル酸キナーゼである請求項1に記載の方法に使用され、前記(b)(i)のアデニル酸キナーゼ不活性化処理及び前記(b)(ii)の血液細胞溶解処理を一段階で実施するべく血液サンプルを収容するための反応容器であって、
前記工程(b)(i)においてアデニル酸キナーゼを不活性化するための試薬であるプロテアーゼ;
前記工程(b)(ii)において血液細胞を溶解するための試薬であるサポニン;
ADP;
ルシフェリン;及び
ルシフェラーゼ
を含んでなる反応容器。
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GB0508981.8 | 2005-05-03 | ||
GBGB0508981.8A GB0508981D0 (en) | 2005-05-03 | 2005-05-03 | Distinguishing cells in a sample |
PCT/GB2006/001618 WO2006117557A2 (en) | 2005-05-03 | 2006-05-03 | Distinguishing cells in a sample by inactivating extracellular enzyme before releasing intracellular enzyme |
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JP2013229488A Pending JP2014042526A (ja) | 2005-05-03 | 2013-11-05 | 細胞内酵素放出前の細胞外酵素不活性化を用いたサンプル中の細胞識別 |
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JP2014042526A (ja) * | 2005-05-03 | 2014-03-13 | 3M Innovative Properties Co | 細胞内酵素放出前の細胞外酵素不活性化を用いたサンプル中の細胞識別 |
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EP2333105A1 (en) * | 2009-12-08 | 2011-06-15 | Koninklijke Philips Electronics N.V. | Selective lysis of cells |
EP3617320B1 (en) * | 2010-04-16 | 2022-01-26 | Momentum Bioscience Limited | Methods for measuring enzyme activity useful in determining cell viability in non-purified samples |
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EP2836610A4 (en) * | 2012-04-12 | 2015-12-30 | Zeus Scientific Inc | METHOD FOR MEASURING POLYMERASE ACTIVITY, USEFUL FOR SENSITIVE QUANTITATIVE MEASUREMENTS OF ANY POLYMERASE EXTENSION ACTIVITIES AND FOR DETERMINING THE PRESENCE OF LIVABLE CELLS |
US8975060B2 (en) * | 2012-11-30 | 2015-03-10 | Qvella Corporation | Method for pretreatment of microbial samples |
US10662420B2 (en) | 2012-11-30 | 2020-05-26 | Qvella Corporation | Apparatus and method for extracting microbial cells |
WO2016044621A1 (en) * | 2014-09-17 | 2016-03-24 | Helixbind, Inc. | Methods and devices for detecting and identifying microorganisms |
JP7073328B2 (ja) | 2016-03-14 | 2022-05-23 | ヘリックスバインド・インコーポレイテッド | 集積流体デバイスと関連する方法 |
AU2019378011A1 (en) * | 2018-11-16 | 2021-06-03 | SeLux Diagnostics, Inc. | System, method and interface for parallel processing of antimicrobial susceptibility tests using different samples |
WO2020202113A1 (en) * | 2019-04-05 | 2020-10-08 | Thermo Fisher Scientific Oy | Mass spectrometric methods and compositions for identifying microbes from blood by reducing confounding proteins |
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IL81822A0 (en) * | 1987-03-08 | 1987-10-20 | Yissum Res Dev Co | Method and kit for testing for the presence of catalase in milk |
JP3200134B2 (ja) * | 1992-02-24 | 2001-08-20 | 湧永製薬株式会社 | マラリア原虫の検出 |
GB9301118D0 (en) | 1993-01-21 | 1993-03-10 | Secr Defence | Enzyme linked assays |
WO1996002665A1 (en) | 1994-07-13 | 1996-02-01 | The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland | Microbiological test method and reagents |
US5843608A (en) | 1995-06-08 | 1998-12-01 | Coulter International Corp. | Reagent and method for differential determination of leukocytes in blood |
GB9902659D0 (en) * | 1999-02-05 | 1999-03-31 | Microbiological Res Authority | Assay with reduced background |
GB9911095D0 (en) * | 1999-05-13 | 1999-07-14 | Secr Defence | Microbiological test method and reagents |
WO2002092843A1 (fr) * | 2001-05-16 | 2002-11-21 | Kikkoman Corporation | Technique de comptage de cellules microbiennes |
GB0308199D0 (en) | 2003-04-09 | 2003-05-14 | Acolyte Biomedica Ltd | Combined microbiology system |
GB0508981D0 (en) * | 2005-05-03 | 2005-06-08 | Acolyte Biomedica Ltd | Distinguishing cells in a sample |
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2005
- 2005-05-03 GB GBGB0508981.8A patent/GB0508981D0/en not_active Ceased
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2006
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- 2006-05-03 MX MX2007013662A patent/MX2007013662A/es active IP Right Grant
- 2006-05-03 AU AU2006242988A patent/AU2006242988A1/en not_active Abandoned
- 2006-05-03 WO PCT/GB2006/001618 patent/WO2006117557A2/en active Application Filing
- 2006-05-03 BR BRPI0611655-8A patent/BRPI0611655A2/pt not_active IP Right Cessation
- 2006-05-03 KR KR1020077028198A patent/KR101315007B1/ko active IP Right Grant
- 2006-05-03 JP JP2008509502A patent/JP5475278B2/ja not_active Expired - Fee Related
- 2006-05-03 EP EP06726995.1A patent/EP1877571B1/en active Active
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2007
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2013
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014042526A (ja) * | 2005-05-03 | 2014-03-13 | 3M Innovative Properties Co | 細胞内酵素放出前の細胞外酵素不活性化を用いたサンプル中の細胞識別 |
Also Published As
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CN101198705A (zh) | 2008-06-11 |
EP1877571B1 (en) | 2015-01-14 |
GB0508981D0 (en) | 2005-06-08 |
WO2006117557A3 (en) | 2007-01-11 |
CN101198705B (zh) | 2013-03-27 |
KR20080056112A (ko) | 2008-06-20 |
MX2007013662A (es) | 2008-04-08 |
JP2008539712A (ja) | 2008-11-20 |
BRPI0611655A2 (pt) | 2011-08-09 |
KR101315007B1 (ko) | 2013-10-04 |
AU2006242988A1 (en) | 2006-11-09 |
US8349583B2 (en) | 2013-01-08 |
JP2014042526A (ja) | 2014-03-13 |
EP1877571A2 (en) | 2008-01-16 |
WO2006117557A2 (en) | 2006-11-09 |
ZA200709442B (en) | 2009-04-29 |
CA2607058A1 (en) | 2006-11-09 |
US20090275063A1 (en) | 2009-11-05 |
US20130130290A1 (en) | 2013-05-23 |
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