WO1999029905B1 - Methode de detection de l'apoptose a l'aide d'oligonucleotides marques par fret - Google Patents
Methode de detection de l'apoptose a l'aide d'oligonucleotides marques par fretInfo
- Publication number
- WO1999029905B1 WO1999029905B1 PCT/US1998/026432 US9826432W WO9929905B1 WO 1999029905 B1 WO1999029905 B1 WO 1999029905B1 US 9826432 W US9826432 W US 9826432W WO 9929905 B1 WO9929905 B1 WO 9929905B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- oligonucleotide
- nucleotide sequence
- moiety
- detectable
- energy
- Prior art date
Links
- 108091034117 Oligonucleotide Proteins 0.000 title claims abstract 20
- 238000000034 method Methods 0.000 title abstract 5
- 230000006907 apoptotic process Effects 0.000 title abstract 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 title 1
- 238000000137 annealing Methods 0.000 claims abstract 4
- 102000003960 Ligases Human genes 0.000 claims abstract 3
- 108090000364 Ligases Proteins 0.000 claims abstract 3
- 239000002773 nucleotide Substances 0.000 claims 20
- 125000003729 nucleotide group Chemical group 0.000 claims 20
- 150000007523 nucleic acids Chemical group 0.000 abstract 2
- 230000003321 amplification Effects 0.000 abstract 1
- 238000006555 catalytic reaction Methods 0.000 abstract 1
- 230000000295 complement effect Effects 0.000 abstract 1
- 230000001404 mediated effect Effects 0.000 abstract 1
- 238000003199 nucleic acid amplification method Methods 0.000 abstract 1
- 238000003752 polymerase chain reaction Methods 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6862—Ligase chain reaction [LCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
L'invention concerne des méthodes d'amplification en chaîne par polymérase induite par ligase (LM-PCR) utilisées pour détecter des fragments cibles d'acides nucléiques à double brin tels que ceux qui sont générés lors du processus apoptosique. Selon ces méthodes, un oligonucléotide détectable est incorporé dans la cible. Cet oligonucléotide détectable renferme la fraction donneur et/ou la fraction receveur d'une paire de transfert d'énergie moléculaire (MET). Un exemple de paire MET est une paire de transfert par résonance d'énergie de fluorescence (FRET) constituée d'une fraction fluorophore (donneur) et d'une fraction extinctrice de luminescence (receveur). La fraction donneur de la paire MET émet de l'énergie détectable telle que de la lumière seulement lorsque l'oligonucléotide détectable est incorporé dans la cible. Selon ces méthodes, un oligonucléotide lieur-amorce est attaché à un oligonucléotide d'aide à la liaison, ou un oligonucléotide lieur-amorce renfermant une séquence d'aide à la liaison est lié à l'extrémité 5' de chacun des brins d'un fragment d'acides nucléiques à double brin renfermant soit une extrémité franche soit un surplomb terminal. Après cette étape de liaison, un oligonucléotide détectable capable d'être attaché au complément de l'oligonuléotide lieur-amorce est incorporé dans la cible par réactions catalysées par polymérase. Selon un autre mode de réalisation, l'oligonucléotide lieur-amorce est également un oligonucléotide détectable. Eventuellement, la cible marquée par l'oligonucléotide détectable est ensuite amplifiée, l'oligonucléotide détectable étant incorporé au produit d'amplification. La cible est détectée par détection de l'énergie émise par le groupe donneur de l'oligonucléotide détectable.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2000524476A JP2001526052A (ja) | 1997-12-12 | 1998-12-11 | Ｆｒｅｔ標識オリゴヌクレオチドを用いたアポトーシスを検出する方法 |
| CA002314225A CA2314225A1 (fr) | 1997-12-12 | 1998-12-11 | Methode de detection de l'apoptose a l'aide d'oligonucleotides marques par fret |
| EP98963108A EP1036201A2 (fr) | 1997-12-12 | 1998-12-11 | Methode de detection de l'apoptose a l'aide d'oligonucleotides marques par fret |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US6943497P | 1997-12-12 | 1997-12-12 | |
| US60/069,434 | 1997-12-12 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO1999029905A2 WO1999029905A2 (fr) | 1999-06-17 |
| WO1999029905A3 WO1999029905A3 (fr) | 1999-08-26 |
| WO1999029905B1 true WO1999029905B1 (fr) | 1999-10-21 |
Family
ID=22088952
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US1998/026432 WO1999029905A2 (fr) | 1997-12-12 | 1998-12-11 | Methode de detection de l'apoptose a l'aide d'oligonucleotides marques par fret |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP1036201A2 (fr) |
| JP (1) | JP2001526052A (fr) |
| CA (1) | CA2314225A1 (fr) |
| WO (1) | WO1999029905A2 (fr) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10048009A1 (de) * | 2000-09-26 | 2002-04-11 | Cytonet Gmbh & Co Kg | Verfahren zum Nachweis von Pilzinfektionen |
| JP2004530410A (ja) * | 2000-10-24 | 2004-10-07 | インタージェン カンパニー | 特異的ヌクレオチド配列の検出 |
| CN1556224B (zh) * | 2004-01-08 | 2012-08-22 | 湖南大学 | 核酸片段检测系统及其在核酸分析及相关酶研究中的应用 |
| WO2007140417A2 (fr) * | 2006-05-31 | 2007-12-06 | Sequenom, Inc. | Procédés et compositions destinés à l'extraction et l'amplification d'un acide nucléique à partir d'un échantillon |
| EP3514244B1 (fr) | 2009-04-03 | 2021-07-07 | Sequenom, Inc. | Procédés de préparation d'acides nucléiques |
| US9957393B2 (en) | 2015-03-30 | 2018-05-01 | Enzo Biochem, Inc. | Monoazo dyes with cyclic amine as fluorescence quenchers |
| CN109182479A (zh) * | 2018-07-27 | 2019-01-11 | 广州医科大学附属第医院 | 一种实时荧光定量pcr通用型荧光探针及其应用 |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5843658A (en) * | 1995-03-10 | 1998-12-01 | Hamamatsu Photonics K.K. | Method of measuring oligonucleotide decomposing activity |
| DE69620766T2 (de) * | 1995-08-11 | 2004-11-18 | Novozymes A/S | Verfahren zur herstellung von polypeptidabkömmlingen |
| WO1997022719A1 (fr) * | 1995-12-18 | 1997-06-26 | Washington University | Procede d'analyse d'acide nucleique par transfert des energies de resonance par fluorescence |
| WO1997039008A1 (fr) * | 1996-04-12 | 1997-10-23 | The Public Health Research Institute Of The City Of New York, Inc. | Sondes, trousses et dosages de detection |
| US6117635A (en) * | 1996-07-16 | 2000-09-12 | Intergen Company | Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon |
-
1998
- 1998-12-11 JP JP2000524476A patent/JP2001526052A/ja active Pending
- 1998-12-11 CA CA002314225A patent/CA2314225A1/fr not_active Abandoned
- 1998-12-11 EP EP98963108A patent/EP1036201A2/fr not_active Withdrawn
- 1998-12-11 WO PCT/US1998/026432 patent/WO1999029905A2/fr not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| WO1999029905A3 (fr) | 1999-08-26 |
| CA2314225A1 (fr) | 1999-06-17 |
| JP2001526052A (ja) | 2001-12-18 |
| EP1036201A2 (fr) | 2000-09-20 |
| WO1999029905A2 (fr) | 1999-06-17 |
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