US5082830A
Expired / Lapsed
US
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End labeled nucleotide probe
Assignee: Enzo Life Sciences Inc
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Filing: 1988-02-26
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Granted: 1992-01-21
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Expires: 2008-02-26
Abstract
Abstract An oligo- or polynucleotide having at least one biotin directly or indirectly attached to each of the 5' and 3' end nucleotides thereof and a nucleic acid hybridization assay system for use therewith. The biotins are optionally attached through linkage groups that do not interfere with hybridization. The biotins can further serve to attach a biotinylated polymer.
Inventors
- 👤 Christine L. Brakel
- 👤 Alan F. Cook
- 👤 Edmund Vuocolo
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 13:27
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6832; C07; C07H; C07H21/00
Claims Analysis
Total Claims: 35
Claim 1
Independent Claim
24 words
An oligo- or polynucleotide having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' end nucleotides thereof.
Claim 2
Dependent Claim
24 words
An oligo- or polynucleotide having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' end nucleotides thereof.
Claim 3
Dependent Claim
17 words
The oligo- or polynucleotide of claim 1 wherein said non-radioactive moiety comprises biotin or a biotin analogue.
Claim 4
Dependent Claim
25 words
The oligo- or polynucleotide of claim 2 comprising at least two molecules of said biotin or biotin analogue attached to at least one end thereof.
Claim 5
Dependent Claim
28 words
The oligo- or polynucleotide of claim 3 comprising from about two to about eighteen molecules of said biotin or biotin analogue attached to at least one end thereof.
Claim 6
Dependent Claim
23 words
The oligo- or polynucleotide of claim 2 comprising at least two molecules of said biotin or biotin analogue attached at each end thereof.
Claim 7
Dependent Claim
25 words
The oligo- or polynucleotide of claim 5 comprising from about two to about eighteen molecules of said biotin or biotin analogue at each end thereof.
Claim 8
Dependent Claim
29 words
The oligo- or polynucleotide of claim 2 wherein at least one molecule of said biotin or biotin analogue is attached to the terminal nucleotide through a non-interfering linkage group.
Claim 9
Dependent Claim
13 words
The oligo- or polynucleotide of claim 7 wherein said biotin attachment comprises biotin-11-dUMP.
Claim 10
Dependent Claim
13 words
The oligo- or polynucleotide of claim 7 wherein said biotin attachment comprises biotin-11-allylamine-dUMP.
Claim 11
Dependent Claim
21 words
The oligo- or polynucleotide of claim 2 wherein at least one of said end nucleotides is attached to a biotinylated polymer.
Claim 12
Dependent Claim
15 words
The oligo- or polynucleotide of claim 10 wherein said biotinylated polymer comprises poly biotinylated dextran.
Claim 13
Dependent Claim
34 words
An oligo- or polynucleotide having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' terminal nucleotides external to a target hybridization region of said oligo- or polynucleotide.
Claim 14
Dependent Claim
34 words
An oligo- or polynucleotide having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' terminal nucleotides external to a target hybridization region of said oligo- or polynucleotide.
Claim 15
Dependent Claim
17 words
The oligo- or polynucleotide of claim 12 wherein said non-radioactive moiety comprises biotin or a biotin analogue.
Claim 16
Dependent Claim
22 words
A nucleic acid hybridization assay composition comprising an oligo- or polynucleotide of claims 1 or 12, and a preformed detectable molecular complex.
Claim 17
Dependent Claim
33 words
A nucleic acid hybridization assay composition comprising an oligo- or polynucleotide of claims 2, 3, 4, 5, 6, 7, 8, 9, 11 or 13, and a preformed avidin or streptavidin detectable molecular complex.
Claim 18
Dependent Claim
29 words
The composition of claim 15 wherein said detectable molecular complex is selected from a fluor, chromogen, enzyme, phosphor, electron dense reagent, or a combination of any of the foregoing.
Claim 19
Dependent Claim
28 words
The composition of claim 16 wherein said enzyme is selected from peroxidase, alkaline phosphatase, acid phosphatase, beta-galactosidase, and glucose oxidase, or a combination of any of the foregoing.
Claim 20
Independent Claim
89 words
A method for detecting a target nucleic acid sequence in a sample comprising: rendering the nucleic acid in said sample in single-stranded form; contacting said single-stranded nucleic acid under hybridizing conditions with (i) an oligo- or polynucleotide probe having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' end nucleotides thereof, said probe being capable of hybridizing to said target nucleic acid sequence, and (ii) a preformed detectable molecular complex; and detecting any hybridized complexes, thereby detecting the target nucleic acid sequence.
Claim 21
Independent Claim
89 words
A method for detecting a target nucleic acid sequence in a sample comprising: rendering the nucleic acid in said sample in single-stranded form; contacting said single-stranded nucleic acid under hybridizing conditions with (i) an oligo- or polynucleotide probe having at least one non-radioactive moiety directly or indirectly attached to each of the 5' and 3' end nucleotides thereof, said probe being capable of hybridizing to said target nucleic acid sequence, and (ii) a preformed detectable molecular complex; and detecting any hybridized complexes, thereby detecting the target nucleic acid sequence.
Claim 22
Dependent Claim
28 words
The method of claim 18 wherein the attachment of said non-radioactive moiety to the oligo- or polynucleotide probe is external to a target hybridization region of said probe.
Claim 23
Dependent Claim
21 words
The method of claim 18 wherein said non-radioactive moiety of the oligo- or polynucleotide probe comprises biotin or a biotin analogue.
Claim 24
Dependent Claim
29 words
The method of claim 20 wherein said oligo- or polynucleotide probe comprises at least two molecules of said biotin or biotin analogue attached to at least one end thereof.
Claim 25
Dependent Claim
29 words
The method of claim 21 wherein said probe comprises from about two to about eighteen molecules of said biotin or biotin analogue attached to at least one end thereof.
Claim 26
Dependent Claim
26 words
The method of claim 20 wherein said oligo- or polynucleotide probe comprises at least two molecules of said biotin or biotin analogue at each end thereof.
Claim 27
Dependent Claim
26 words
The method of claim 23 wherein said probe comprises from about two to about eighteen molecules of said biotin or biotin analogue at each end thereof.
Claim 28
Dependent Claim
30 words
The method of claim 20 wherein at least one of said biotin or biotin analogue molecules in said probe is attached to the terminal nucleotide through a non-interfering linkage group.
Claim 29
Dependent Claim
11 words
The method of claim 25 wherein said biotin attachment comprises biotin-11-dUMP.
Claim 30
Dependent Claim
11 words
The method of claim 25 wherein said biotin attachment comprises biotin-11-allylamine-dUMP.
Claim 31
Dependent Claim
22 words
The method of claim 18 wherein at least one of said end nucleotides in the probe is attached to a biotinylated polymer.
Claim 32
Dependent Claim
13 words
The method of claim 28 wherein said biotinylated polymer comprises poly biotinylated dextran.
Claim 33
Dependent Claim
14 words
The method of claim 20 wherein said detectable molecular complex comprises avidin or streptavidin.
Claim 34
Independent Claim
31 words
The method of claims 18 or 30 wherein said detectable molecular complex is selected from a fluor, chromogen, enzyme, phosphor, electron dense reagent, or a combination of any of the foregoing.
Claim 35
Dependent Claim
28 words
The method of claim 31 wherein said enzyme is selected from peroxidase, alkaline phosphatase, acid phosphatase, beta-galactosidase, and glucose oxidase, or a combination of any of the foregoing.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1995-01-20 | 1995-07-22 | 1996-01-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-01-20 | 1999-07-22 | 2000-01-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-01-20 | 2003-07-22 | 2004-01-21 | $7700.00 | Window Closed / Lapsed |