WO1999029905A3 - Method for detecting apoptosis using fret labeled oligonucleotides - Google Patents

Method for detecting apoptosis using fret labeled oligonucleotides Download PDF

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Publication number
WO1999029905A3
WO1999029905A3 PCT/US1998/026432 US9826432W WO9929905A3 WO 1999029905 A3 WO1999029905 A3 WO 1999029905A3 US 9826432 W US9826432 W US 9826432W WO 9929905 A3 WO9929905 A3 WO 9929905A3
Authority
WO
WIPO (PCT)
Prior art keywords
oligonucleotide
detectable
target
moiety
linker
Prior art date
Application number
PCT/US1998/026432
Other languages
French (fr)
Other versions
WO1999029905B1 (en
WO1999029905A2 (en
Inventor
William M James
Irina A Nazarenko
Original Assignee
Intergen Company
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Intergen Company filed Critical Intergen Company
Priority to EP98963108A priority Critical patent/EP1036201A2/en
Priority to CA002314225A priority patent/CA2314225A1/en
Priority to JP2000524476A priority patent/JP2001526052A/en
Publication of WO1999029905A2 publication Critical patent/WO1999029905A2/en
Publication of WO1999029905A3 publication Critical patent/WO1999029905A3/en
Publication of WO1999029905B1 publication Critical patent/WO1999029905B1/en

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    • 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]

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Ligase-mediated polymerase chain reaction (LM-PCR) methods are used to detect target double-stranded nucleic acid fragments such as those generated during the process of apoptosis. In these methods, a detectable oligonucleotide is incorporated into the target. This detectable oligonucleotide contains the donor moiety and/or the acceptor moiety of a molecular energy transfer (MET) pair. An example of a MET pair is a fluorescence energy resonance transfer (FRET) pair consisting of a fluorophore (donor) moiety and a quencher (acceptor) moiety. The donor moiety of the MET pair emits detectable energy such as light only when the detectable oligonucleotide is incorporated into the target. In these methods, a linker-primer oligonucleotide annealed to a ligation-aid oligonucleotide, or a linker-primer oligonucleotide containing a ligation-aid sequence, is ligated to the 5' end of each strand of a double-stranded nucleic acid fragment containing either a blunt end or a terminal overhang. After this ligation step, a detectable oligonucleotide capable of annealing to the complement of the linker-primer oligonucleotide is incorporated into the target by polymerase-catalyzed reactions. Alternatively, the linker-primer oligonucleotide is also a detectable oligonucleotide. Optionally, the target labeled by the detectable oligonucleotide is subsequently amplified, wherein the detectable oligonucleotide is incorporated into the amplification product. The target is detected by detecting the energy emitted by the donor moiety of the detectable oligonucleotide.
PCT/US1998/026432 1997-12-12 1998-12-11 Method for detecting apoptosis using fret labeled oligonucleotides WO1999029905A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP98963108A EP1036201A2 (en) 1997-12-12 1998-12-11 Method for detecting apoptosis using fret labeled oligonucleotides
CA002314225A CA2314225A1 (en) 1997-12-12 1998-12-11 Method for detecting apoptosis using fret labeled oligonucleotides
JP2000524476A JP2001526052A (en) 1997-12-12 1998-12-11 Method for detecting apoptosis using FRET-labeled oligonucleotide

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 (en) 1999-06-17
WO1999029905A3 true WO1999029905A3 (en) 1999-08-26
WO1999029905B1 WO1999029905B1 (en) 1999-10-21

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1998/026432 WO1999029905A2 (en) 1997-12-12 1998-12-11 Method for detecting apoptosis using fret labeled oligonucleotides

Country Status (4)

Country Link
EP (1) EP1036201A2 (en)
JP (1) JP2001526052A (en)
CA (1) CA2314225A1 (en)
WO (1) WO1999029905A2 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10048009A1 (en) * 2000-09-26 2002-04-11 Cytonet Gmbh & Co Kg Method of detecting fungal infections
JP2004530410A (en) * 2000-10-24 2004-10-07 インタージェン カンパニー Detection of specific nucleotide sequences
CN1556224B (en) * 2004-01-08 2012-08-22 湖南大学 Nucleic acid fragment detection system and its application in nucleic acid analysis and related enzyme study
US8679741B2 (en) 2006-05-31 2014-03-25 Sequenom, Inc. Methods and compositions for the extraction and amplification of nucleic acid from a sample
EP3514244B1 (en) 2009-04-03 2021-07-07 Sequenom, Inc. Nucleic acid preparation methods
US9957393B2 (en) 2015-03-30 2018-05-01 Enzo Biochem, Inc. Monoazo dyes with cyclic amine as fluorescence quenchers
CN109182479A (en) * 2018-07-27 2019-01-11 广州医科大学附属第医院 A kind of real-time fluorescence quantitative PCR Universal fluorescence probe and its application

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0731178A2 (en) * 1995-03-10 1996-09-11 Hamamatsu Photonics K.K. Method of measuring oligonucleotide decomposition
WO1997007205A1 (en) * 1995-08-11 1997-02-27 Novo Nordisk A/S Method for preparing polypeptide variants
WO1997022719A1 (en) * 1995-12-18 1997-06-26 Washington University Method for nucleic acid analysis using fluorescence resonance energy transfer
WO1997039008A1 (en) * 1996-04-12 1997-10-23 The Public Health Research Institute Of The City Of New York, Inc. Detection probes, kits and assays
WO1998002449A1 (en) * 1996-07-16 1998-01-22 Oncor, Inc. Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0731178A2 (en) * 1995-03-10 1996-09-11 Hamamatsu Photonics K.K. Method of measuring oligonucleotide decomposition
WO1997007205A1 (en) * 1995-08-11 1997-02-27 Novo Nordisk A/S Method for preparing polypeptide variants
WO1997022719A1 (en) * 1995-12-18 1997-06-26 Washington University Method for nucleic acid analysis using fluorescence resonance energy transfer
WO1997039008A1 (en) * 1996-04-12 1997-10-23 The Public Health Research Institute Of The City Of New York, Inc. Detection probes, kits and assays
WO1998002449A1 (en) * 1996-07-16 1998-01-22 Oncor, Inc. Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
CANTOR: "LIGHTING UP HYBRIDIZATION", BIO/TECHNOLOGY, vol. 14, 1 March 1996 (1996-03-01), pages 247, XP002094958 *
HIYOSHI M ET AL: "ASSAY OF DNA DENATURATION BY POLYMERASE CHAIN REACTION-DRIVEN FLUORESCENT LABEL INCORPORATION AND FLUORESCENCE RESONANCE ENERGY TRANSFER", ANALYTICAL BIOCHEMISTRY, vol. 221, no. 2, 1 September 1994 (1994-09-01), pages 306 - 311, XP000460389 *
JAMES W AND NAZARENKO I: "Fluorescence quantitation technique for apoptotic DNA fragmentation", PROCEEDINGS OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH ANNUAL MEETING, vol. 39, 1998, pages 580, XP002104356 *
JU J ET AL: "ENERGY TRANSFER PRIMERS: A NEW FLUORESCENCE LABELING PARADIGM FOR DNA SEQUENCING AND ANALYSIS", NATURE MEDICINE, vol. 2, no. 2, 1 February 1996 (1996-02-01), pages 246 - 249, XP002063932 *
NAZARENKO ET AL: "A CLOSED TUBE FORMAT FOR AMPLIFICATION AND DETECTION OF DNA BASED ENERGY TRANSFER", NUCLEIC ACIDS RESEARCH, vol. 25, no. 12, 1 January 1997 (1997-01-01), pages 2516 - 2521, XP002094959 *
STALEY K ET AL.: "Apoptotic DNA fragmentation is detected by a semi-quantitative ligation-mediated PCR of blunt DNA ends", CELL DEATH AND DFFERENTIATION, vol. 4, 1997, pages 66 - 75, XP002104355 *
WU D Y ET AL: "THE LIGATION AMPLIFICATION REACTION (LAR)- AMPLIFICATION OF SPECIFIC DNA SEQUENCES USING SEQUENTIAL ROUNDS OF TEMPLE-DEPENDENT LIGATION", GENOMICS, vol. 4, 1 January 1989 (1989-01-01), pages 560 - 569, XP000318979 *

Also Published As

Publication number Publication date
JP2001526052A (en) 2001-12-18
EP1036201A2 (en) 2000-09-20
WO1999029905B1 (en) 1999-10-21
CA2314225A1 (en) 1999-06-17
WO1999029905A2 (en) 1999-06-17

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