US5759781A
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Multiparametric fluorescence in situ hybridization
Assignee: Qiagen GmbH
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Filing: 1996-05-01
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Granted: 1998-06-02
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Expires: 2016-05-01
Abstract
Abstract The invention relates to a set of combinatorially labeled oligonucleotide probes each member thereof: (i) having a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype, and to the use of such molecules to identify the individual autosomal and sex chromosomes of a human karyotype.
Inventors
- 👤 David C. Ward
- 👤 Michael Speicher
- 👤 Stephen Gwynn Ballard
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 02:53
CPC Classifications
C; C07; C07H; C07H21/00; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6841
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
81 words
A set of combinatorially labeled oligonucleotide probes, each member thereof comprising a plurality of an oligonucleotide, said oligonucleotide: (i) being linked or coupled to a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal or sex chromosome of said human karyotype to at least one member.
Claim 2
Dependent Claim
81 words
A set of combinatorially labeled oligonucleotide probes, each member thereof comprising a plurality of an oligonucleotide, said oligonucleotide: (i) being linked or coupled to a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal or sex chromosome of said human karyotype to at least one member.
Claim 3
Dependent Claim
34 words
The set of combinatorially labeled oligonucleotide probes of claim 1, wherein said set contains at least one member probe specific for a sub-chromosomal fragment of an autosomal or sex chromosome of said human karyotype.
Claim 4
Dependent Claim
23 words
The set of combinatorially labeled oligonucleotide probes of claim 1, wherein said probes contain nucleotide residues that are labeled with a biotin moiety.
Claim 5
Dependent Claim
21 words
The set of combinatorially labeled oligonucleotide probes of claim 3, wherein said probes are additionally labeled with a labeled biotin-binding ligand.
Claim 6
Dependent Claim
29 words
The set of combinatorially labeled oligonucleotide probes of claim 4, wherein at least one of said probes is labeled with a biotin-binding ligand that comprises one or more fluorophores.
Claim 7
Dependent Claim
33 words
The set of combinatorially labeled oligonucleotide probes of claim 5, wherein at least one of said fluorophores is selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.
Claim 8
Dependent Claim
29 words
The set of combinatorially labeled oligonucleotide probes of claim 5, wherein at least one of said probes is labeled with a biotin-binding ligand that comprises more than one fluorophore.
Claim 9
Dependent Claim
40 words
The set of combinatorially labeled oligonucleotide probes of claim 7, wherein one of the fluorophores of said biotin-binding ligand that comprises more than one fluorophore is selected from the group consisting of fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.
Claim 10
Dependent Claim
40 words
The set of combinatorially labeled oligonucleotide probes of claim 8, wherein all of the fluorophores of said biotin-binding ligand that comprises more than one fluorophores are selected from the group consisting of fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.
Claim 11
Dependent Claim
60 words
The set of combinatorially labeled oligonucleotide probes of claim 5, wherein at least one member of said set is labeled with more than one fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy 3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 12
Dependent Claim
59 words
The set of combinatorially labeled oligonucleotide probes of claim 10, wherein at least one member of said set is labeled with two fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 13
Dependent Claim
59 words
The set of combinatorially labeled oligonucleotide probes of claim 10, wherein at least one member of said set is labeled with three fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 14
Independent Claim
233 words
A method of simultaneously identifying and distinguishing the individual autosomal and sex chromosomes of a human karyotype which comprises the steps: (a) contacting a preparation of said chromosomes, in single-stranded form, under conditions sufficient to permit nucleic acid oligonucleotide probes, each member thereof comprising a plurality of an oligonucleotide, said oligonucleotide: (i) being linked or coupled to a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal or sex chromosome of said human karyotype to at least one member; wherein said contacting thereby causes at least one of each autosomal or sex chromosome of said preparation to become hybridized to at least one member of said set of probes; (b) for each chromosome of said preparation hybridized to a member of said set probes, detecting and identifying the predetermined label of said hybridized probe member and correlating the identity of the label of said hybridized probe member with the identity of the autosomal or sex chromosome of said human karyotype with which said hybridized probe member specifically hybridizes, to thereby identify the chromosome hybridized to said member; and (c) repeating step (b) until each autosomal and sex chromosome of said human karyotype has been identified in said preparation.
Claim 15
Independent Claim
233 words
A method of simultaneously identifying and distinguishing the individual autosomal and sex chromosomes of a human karyotype which comprises the steps: (a) contacting a preparation of said chromosomes, in single-stranded form, under conditions sufficient to permit nucleic acid oligonucleotide probes, each member thereof comprising a plurality of an oligonucleotide, said oligonucleotide: (i) being linked or coupled to a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal or sex chromosome of said human karyotype to at least one member; wherein said contacting thereby causes at least one of each autosomal or sex chromosome of said preparation to become hybridized to at least one member of said set of probes; (b) for each chromosome of said preparation hybridized to a member of said set probes, detecting and identifying the predetermined label of said hybridized probe member and correlating the identity of the label of said hybridized probe member with the identity of the autosomal or sex chromosome of said human karyotype with which said hybridized probe member specifically hybridizes, to thereby identify the chromosome hybridized to said member; and (c) repeating step (b) until each autosomal and sex chromosome of said human karyotype has been identified in said preparation.
Claim 16
Dependent Claim
32 words
The method of claim 13, wherein said set of oligonucleotide probes contains at least one member probe specific for a sub-chromosomal fragment of an autosomal or sex chromosome of said human karyotype.
Claim 17
Dependent Claim
18 words
The method of claim 13, wherein said probes contain nucleotide residues that are labeled with a biotin moiety.
Claim 18
Dependent Claim
16 words
The method of claim 15, wherein said probes are additionally labeled with a labeled biotin-binding ligand.
Claim 19
Dependent Claim
24 words
The method of claim 16, wherein at least one of said probes is labeled with a biotin-binding ligand that comprises one or more fluorophores.
Claim 20
Dependent Claim
28 words
The method of claim 17, wherein at least one of said fluorophores is selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7.
Claim 21
Dependent Claim
24 words
The method of claim 17, wherein at least one of said probes is labeled with a biotin-binding ligand that comprises more than one fluorophore.
Claim 22
Dependent Claim
35 words
The method of claim 19, wherein one of the fluorophores of said biotin-binding ligand that comprises more than one fluorophore is selected from the group consisting of fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5, and Cy7.
Claim 23
Dependent Claim
35 words
The method of claim 20, wherein all of the fluorophores of said biotin-binding ligand that comprises more than one fluorophore are selected from the group consisting of fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5, and Cy7.
Claim 24
Dependent Claim
54 words
The method of claim 17, wherein at least one member of said set is labeled with more than one fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 25
Dependent Claim
54 words
The method of claim 22, wherein at least one member of said set is labeled with two fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 26
Dependent Claim
54 words
The method of claim 22, wherein at least one member of said set is labeled with three fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 27
Dependent Claim
24 words
The method of claim 13, wherein an optical comb is employed in step (b) to detect the predetermined label of said hybridized probe member.
Claim 28
Dependent Claim
24 words
The method of claim 13, wherein an optical filter is employed in step (b) to detect the predetermined label of said hybridized probe member.
Claim 29
Dependent Claim
27 words
The method of claim 26, wherein a plurality of optical filters is sequentially employed in step (b) to detect the predetermined label of said hybridized probe member.
Claim 30
Dependent Claim
54 words
The method of claim 26, wherein at least one member of said set is labeled with more than one fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 31
Dependent Claim
54 words
The method of claim 26, wherein at least one member of said set is labeled with two fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
Claim 32
Dependent Claim
54 words
The method of claim 26, wherein at least one member of said set is labeled with three fluorophores, each fluorophore selected from the group consisting of the fluorophores FITC, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7, and wherein each member of said set is labeled with at least one fluorophore selected from said fluorophore group.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-06-01 | 2001-12-01 | 2002-06-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-06-01 | 2005-12-01 | 2006-06-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-06-01 | 2009-12-01 | 2010-06-02 | $7700.00 | Window Closed / Lapsed |