Patent US4868105A successfully refreshed from Google Patents.
US4868105A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Solution phase nucleic acid sandwich assay
Assignee: Bayer Corp
•
Filing: 1985-12-11
•
Granted: 1989-09-19
•
Expires: 2005-12-11
Abstract
Abstract Methods and compositions are provided for rapid detection of nucleic acid sequences. The method employs two reagent sets. The first set is a labelling set comprising: (1) a first nucleic acid sequence probe having an analyte complementary region and a first recognition sequence region and (2) a labelled sequence complementary to the first recognition sequence region. The second set is a capturing set comprising: (1) a second nucleic acid sequence probe having an analyte complementary region and a second recognition sequence region, (2) a specific binding pair member conjugated to a sequence complementary to the second recognition sequence, and (3) a separating means to which is bound a complementary specific binding pair member. The sample and probes are combined under annealing conditions, followed by addition of the other reagents, separation of the bound label from the supernatant and detection of the label in either phase.
Inventors
- 👤 Mickey Urdea
- 👤 Brian Warner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 11:42
CPC Classifications
C; C07; C07H; C07H21/00; C07H19/00; C07H19/02; C07H19/04; C07H19/06; C07H19/10; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/682; C12Q1/70; C12Q1/701; C12Q1/706; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
415 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing one member of a specific binding receptor-ligand pair, the other, complementary member of said specific binding pair being covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 2
Dependent Claim
415 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing one member of a specific binding receptor-ligand pair, the other, complementary member of said specific binding pair being covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein said solid support comprises a plurality of particles.
Claim 4
Dependent Claim
14 words
The method of claim 1, wherein said label is a fluorescer or an enzyme.
Claim 5
Dependent Claim
15 words
The method of claim 1, wherein said receptor is biotin and said ligand is avidin.
Claim 6
Dependent Claim
394 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 7
Dependent Claim
394 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 8
Dependent Claim
29 words
The method of claim 5, wherein said analyte is initially provided in double-stranded form and, prior to said combining, is denatured in the presence of base at elevated temperature.
Claim 9
Dependent Claim
14 words
The method of claim 5, wherein said solid support comprises a plurality of particles.
Claim 10
Dependent Claim
14 words
The method of claim 5, wherein said label is a fluorescer or an enzyme.
Claim 11
Dependent Claim
414 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing one member of a specific binding hapten-antibody pair, the other, complementary member of said specific binding pair being covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 12
Dependent Claim
414 words
An assay method for detecting a nucleic acid analyte in a sample, employing two sets of reagents, a labelling set and a capturing set, said method comprising: (1) providing members of said labelling set of reagents, comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) providing members of said capturing set of reagents, comprising: (c) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length which is complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte, and wherein each of said sequences C-1 is complementary to physically distinct, nonoverlapping sequences of said analyte; and (d) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing one member of a specific binding hapten-antibody pair, the other, complementary member of said specific binding pair being covalently bound to a solid support; (3) combining in a liquid medium under binding conditions for complementary pairs, said sample containing analyte in single-stranded form, with said labelling and capturing probes, to provide probe-analyte complexes; (4) combining said probe-analyte complexes with said capturing reagent to give support-bound probe-analyte complexes; (5) washing said support to provide said support-bound probe-analyte complexes substantially free of unbound probe and unbound analyte; (6) contacting said support with said labelling reagent to label said support-bound probe-analyte complexes; (7) separating said label into a bound phase and an unbound phase by means of said solid support; and (8) detecting the amount of bound or unbound label as determinative of the presence of said analyte.
Claim 13
Dependent Claim
14 words
The method of claim 9, wherein said solid support comprises a plurality of particles.
Claim 14
Dependent Claim
14 words
The method of claim 9, wherein said label is a fluorescer or an enzyme.
Claim 15
Dependent Claim
29 words
The method of claim 9, wherein said binding of said capturing reagent to said solid support is effected at a pH in the range of about 6 to 9.
Claim 16
Dependent Claim
297 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing a first member of a specific binding receptor-ligand pair; and (c) a solid support, to which is bound a second member of said specific binding pair, complementary to said first member, enabling separation and detection of said analyte.
Claim 17
Dependent Claim
297 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing a first member of a specific binding receptor-ligand pair; and (c) a solid support, to which is bound a second member of said specific binding pair, complementary to said first member, enabling separation and detection of said analyte.
Claim 18
Dependent Claim
15 words
The kit of claim 13, wherein said receptor is biotin and said ligand is avidin.
Claim 19
Dependent Claim
14 words
The kit of claim 13, wherein said solid support comprises a plurality of particles.
Claim 20
Dependent Claim
14 words
The kit of claim 13, wherein said label is a fluorescer or an enzyme.
Claim 21
Dependent Claim
280 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary, region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2; and (c) a solid support enabling separation and detection of said analyte, wherein said capturing reagent is covalently bound to said solid support.
Claim 22
Dependent Claim
280 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region at most about 5 kb in length and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary, region at most about 5 kb in length and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2; and (c) a solid support enabling separation and detection of said analyte, wherein said capturing reagent is covalently bound to said solid support.
Claim 23
Dependent Claim
14 words
The kit of claim 17, wherein said solid support comprises a plurality of particles.
Claim 24
Dependent Claim
297 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region containing at most about 5 kb in total and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region containing at most about 5 kb and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing a first member of a specific binding hapten-antibody pair; and (c) a solid support, to which is bound a second member of said specific binding pair, complementary to said first member, enabling separation and detection of said analyte.
Claim 25
Dependent Claim
297 words
A kit for detecting a nucleic acid analyte in single-stranded form, comprising: (1) members of a labelling set of reagents comprising: (a) a plurality of labelling nucleic acid probes comprising single-stranded polynucleotide chains each having two nucleic acid regions, the first region having a nucleic acid sequence L-1 about 15 to about 100 nucleotides in length and selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region containing at most about 5 kb in total and including a labelling reagent recognition sequence L-2, wherein each of said sequences L-1 is selected so as to be complementary to physically distinct, nonoverlapping sequences of said analyte; and (b) a labelling reagent having a nucleic acid sequence complementary to said labelling reagent recognition sequence L-2 and a label which provides, directly or indirectly, a detectable signal; and (2) members of a capturing set of reagents comprising: (a) a plurality of capturing nucleic acid probes comprising single-stranded polynucleotide chains each having two polynucleotide sequence regions, the first region having a nucleic acid sequence C-1 about 15 to about 100 nucleotides in length and which is selected so as to be complementary to a sequence of said analyte, and the second, noncomplementary region containing at most about 5 kb and including a capturing reagent recognition sequence C-2, wherein said sequences L-1 and C-1 are nonidentical, noncomplementary sequences that are each complementary to physically distinct sequences of said analyte; (b) a capturing reagent having a nucleic acid sequence complementary to said capturing reagent recognition sequence C-2 and containing a first member of a specific binding hapten-antibody pair; and (c) a solid support, to which is bound a second member of said specific binding pair, complementary to said first member, enabling separation and detection of said analyte.
Claim 26
Dependent Claim
14 words
The kit of claim 19, wherein said solid support comprises a plurality of particles.
Claim 27
Dependent Claim
14 words
The kit of claim 19, wherein said label is a fluorescer or an enzyme.
References Cited (Backward Citations)
GB215607A
Cited
WO1983001459A1
Cited
WO1984003285A1
Cited
WO1984003520A1
Cited
EP0123300A2
Cited
EP0128332A1
Cited
EP0139489A2
Cited
US4731325A
Cited
US4716106A
Cited
US4563417A
Cited
GB2169403A
Cited
WO1986006412A1
Cited
US4751177A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-09-18 | 1993-03-20 | 1993-09-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-09-18 | 1997-03-20 | 1997-09-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-09-18 | 2001-03-20 | 2001-09-19 | $7700.00 | Window Closed / Lapsed |