EP0601889A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Nucleic acid probes
Assignee: Maine Medical Center Research Institute
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Filing: 1993-12-10
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Granted: —
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Expires: 2013-12-10
Abstract
Abstract A probe includes (1) a nucleotide sequence which has a segment complementary to the polynucleotide target and is capable of forming one or more imperfect hairpins with at least one of the hairpins including the segment or a part thereof; and (2) at least one donor label moiety and at least one acceptor label moiety covalently attached to the nucleotide sequence so that only when one more imperfect hairpins are formed, one of the donor label moieties and one of the acceptor label moieties are in close proximity to allow resonance energy transfer between them. The label moieties are preferably fluorophores. Also disclosed is a method of using such a probe.
Inventors
- 👤 Bruce C. Bagwell
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-28 22:25
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
103 words
A probe for binding to a polynucleotide target, which probe comprises: a nucleotide sequence having a segment complementary to the polynucleotide target, said nucleotide sequence being capable of forming one or more imperfect hairpins at least one of which includes said segment or a part thereof; and at least one donor label moiety and at least one acceptor label moiety covalently attached to said nucleotide sequence so that only when said one or more imperfect hairpins are formed, said at least one donor label moiety and said at least one acceptor label moiety are in close proximity to allow resonance energy transfer therebetween.
Claim 2
Dependent Claim
24 words
A probe according to Claim 1, wherein said donor label moiety is a donor fluorophore and said acceptor label moiety is an acceptor fluorophore.
Claim 3
Dependent Claim
20 words
A probe according to Claims 1 or 2, wherein said nucleotide sequence is capable of forming only one imperfect hairpin.
Claim 4
Dependent Claim
70 words
A probe according to Claim 3, wherein two donor and acceptor label moieties are covalently attached to said nucleotide sequence, with one attached to an internal nucleotide of said nucleotide sequence and the other attached to another internal nucleotide of said nucleotide sequence so that only when said one imperfect hairpin is formed, said two donor and acceptor label moieties are in close proximity to allow resonance energy transfer therebetween.
Claim 5
Dependent Claim
16 words
A probe according to Claims 3 or 4, wherein said segment contains 22 nucleotides or more.
Claim 6
Dependent Claim
20 words
A probe according to any of Claims 3, 4 or 5, wherein said donor and acceptor label moieties are identical.
Claim 7
Dependent Claim
47 words
A probe according to both Claim 2 and any of Claims 3 to 6, further comprising an additional fluorophore covalently attached to said nucleotide sequence, said additional fluorophore being remote from, and emitting at a wavelength other than the emission wavelengths of, said donor and acceptor fluorophores.
Claim 8
Dependent Claim
20 words
A probe according to Claims 1 or 2, wherein said nucleotide sequence is capable of forming only two imperfect hairpins.
Claim 9
Dependent Claim
87 words
A probe according to Claim 8, wherein three donor and acceptor label moieties are covalently attached to said nucleotide sequence, with one attached to a nucleotide at one end of said nucleotide sequence, another attached to a nucleotide at the other end of said nucleotide sequence, and the third one attached to a nucleotide near the centre of said segment so that only when two imperfect hairpins are formed, said three donor and acceptor label moieties are all in close proximity to allow resonance energy transfer therebetween.
Claim 10
Dependent Claim
16 words
A probe according to Claims 8 or 9, wherein said segment contains 34 nucleotides or more.
Claim 11
Dependent Claim
18 words
A probe according to any of Claims 9 or 10, wherein said donor and acceptor fluorophores are identical.
Claim 12
Dependent Claim
47 words
A probe according to both Claim 2 and any of Claims 8 to 11, further comprising an additional fluorophore covalently attached to said nucleotide sequence, said additional fluorophore being remote from, and emitting at a wavelength other than the emission wavelengths of, said donor and acceptor fluorophores.
Claim 13
Dependent Claim
79 words
A method for detecting a polynucleotide target in a sample, which method comprises: contacting the sample with a probe according to Claim 2, having at least one donor fluorophore and at least one acceptor fluorophore; dissociating double-stranded nucleic acids into single-stranded nucleic acids and disrupting secondary structures; hybridizing said probe with the polynucleotide target; and monitoring fluorescent emission change of said at least one acceptor fluorophore upon irradiation of said at least one donor fluorophore with an excitation light.
Claim 14
Dependent Claim
24 words
The method of Claim 13, wherein said monitoring step is performed by measurement of fluorescent emission intensity of said at least one acceptor fluorophore.
References Cited (Backward Citations)
No citation records indexed.
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