Global patent index - EP 1036201 A2

EP 1036201 A2 2000-09-20 - METHOD FOR DETECTING APOPTOSIS USING FRET LABELED OLIGONUCLEOTIDES

Title (en)

METHOD FOR DETECTING APOPTOSIS USING FRET LABELED OLIGONUCLEOTIDES

Title (de)

METHODE ZUM NACHWEIS VON APOPTOSE UNTER VERWENDUNG VON "FRET" MARKIERTEN OLIGONUKLEOTIDEN

Title (fr)

METHODE DE DETECTION DE L'APOPTOSE A L'AIDE D'OLIGONUCLEOTIDES MARQUES PAR FRET

Publication

EP 1036201 A2 (EN)

Application

EP 98963108 A

Priority

  • US 9826432 W
  • US 6943497 P

Abstract (en)

[origin: WO9929905A2] 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.

IPC 1-7 (main, further and additional classification)

C12Q 1/68

IPC 8 full level (invention and additional information)

C12N 15/09 (2006.01); C12Q 1/68 (2006.01)

CPC (invention and additional information)

C12Q 1/6818 (2013.01); C12Q 1/6862 (2013.01)

Combination set (CPC)

  1. C12Q 1/6818 + C12Q 2531/137 + C12Q 2525/301 + C12Q 2525/197
  2. C12Q 1/6862 + C12Q 2565/101 + C12Q 2525/301 + C12Q 2525/197

Citation (search report)

See references of WO 9929905A3

Designated contracting state (EPC)

AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

EPO simple patent family

WO 9929905 A2 19990617; WO 9929905 A3 19990826; WO 9929905 B1 19991021; CA 2314225 A1 19990617; EP 1036201 A2 20000920; JP 2001526052 A 20011218

INPADOC legal status

2005-01-05 [18D] DEEMED TO BE WITHDRAWN

- Ref Legal Event Code: 18D

- Effective date: 20040702

2000-09-20 [17P] REQUEST FOR EXAMINATION FILED

- Ref Legal Event Code: 17P

- Effective date: 20000712

2000-09-20 [AK] DESIGNATED CONTRACTING STATES:

- Ref Legal Event Code: AK

- Designated State(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE