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US4868104A Expired / Lapsed US ⏱️ 0.0 years remaining

Homogeneous assay for specific polynucleotides

Assignee: Siemens Healthcare Diagnostics GmbH Germany
•
Filing: 1985-09-06
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Granted: 1989-09-19
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Expires: 2005-09-06
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Abstract

Abstract A method for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte is disclosed. The method comprises combining in an assay medium the sample and first and second polynucleotide reagents complementary to the analyte. Each of the first and second reagents hybridize with a different region of the analyte. The first reagent contains means for rendering the first reagent non-covalently polymerizable. The second reagent contains means for rendering the second reagent detectable. The sample and the first and second reagents are combined in the assay medium under conditions for polymerizing the first reagent wherein the second reagent becomes bound to the polymerized first reagent only when the analyte is present in the sample. A determination is then made as to whether the second reagent has become bound to the polymerized first reagent. The method has broad application for determining the presence of a polynucleotide analyte such as DNA, RNA, the genomes of viruses, bacteria, molds, fungi, and fragments thereof, and the like. Preferred means for rendering the first reagent non-covalently polymerizable includes a repeating oligonucleotide sequence covalently bound to the first reagent.

Inventors

  • 👤 Nurith Kurn
  • 👤 Chander Bahl
  • 👤 Edwin F. Ullman

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-24 22:48
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/705; C12Q1/68; C12Q1/6813; C12Q1/6832; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S436/00; Y10S436/807; Y10S436/808

Claims Analysis

Total Claims: 102
Claim 1
Independent Claim 127 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and first and second polynucleotide reagents complementary to said analyte, wherein each of said first and second reagents hybridizes with a different region of said analyte, said first reagent containing means for rendering said first reagent non-covalently polymerizable and said second reagent containing means for rendering said second reagent detectable, wherein, under conditions for polymerizing said first reagent, said second reagent becomes bound to said polymerized first reagent only when said analyte is present in said sample, (b) non-covalently polymerizing said first reagent, and (c) determining whether said second reagent has become bound to said polymerized first reagent.
Claim 2
Independent Claim 127 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and first and second polynucleotide reagents complementary to said analyte, wherein each of said first and second reagents hybridizes with a different region of said analyte, said first reagent containing means for rendering said first reagent non-covalently polymerizable and said second reagent containing means for rendering said second reagent detectable, wherein, under conditions for polymerizing said first reagent, said second reagent becomes bound to said polymerized first reagent only when said analyte is present in said sample, (b) non-covalently polymerizing said first reagent, and (c) determining whether said second reagent has become bound to said polymerized first reagent.
Claim 3
Dependent Claim 23 words
The method of claim 1 wherein a compound capable of causing the non-covalent polymerization of said first reagent is combined in Step b.
Claim 4
Dependent Claim 11 words
The method of claim 1 wherein said polynucleotide analyte is DNA.
Claim 5
Dependent Claim 11 words
The method of claim 1 wherein said polynucleotide analyte is RNA.
Claim 6
Dependent Claim 28 words
The method of claim 1 wherein Step c said polymerized first reagent is examined for the presence of said second reagent by determining the presence of aggregated particles.
Claim 7
Dependent Claim 25 words
The method of claim 1 wherein said polynucleotide analyte is selected from the group consisting of genomes of viruses, bacteria, and fungi, and fragments thereof.
Claim 8
Dependent Claim 10 words
The method of claim 6 wherein the fungi are molds.
Claim 9
Dependent Claim 57 words
The method of claim 1 wherein said means for rendering said first reagent non-covalently polymerizable includes a first specific binding pair (sbp) member bound to said first reagent and wherein a second sbp member complementary to said first sbp member having at least two specific binding sites for said sbp member is combined with said assay medium.
Claim 10
Dependent Claim 28 words
The method of claim 8 wherein said sbp member and said complementary sbp member are selected from the group consisting of antigen-antibody, avidin-biotin, IgG-protein A, DNA-DNA and DNA-RNA.
Claim 11
Dependent Claim 26 words
The method of claim 1 wherein said means for rendering said first reagent non-covalently polymerizable includes a repeating oligonucleotide sequence covalently bound to said first reagent.
Claim 12
Dependent Claim 19 words
The method of claim 10 wherein said repeating oligonucleotide sequence is a homopolymer having at least 8 nucleotide residues.
Claim 13
Dependent Claim 24 words
The method of claim 10 wherein said repeating oligonucleotide sequence is a heteropolymer at least eight nucleotides in length consisting of two non-complementary nucleotides.
Claim 14
Dependent Claim 59 words
The method of claim 1 wherein said means for rendering said second reagent detectable includes a first specific binding pair (sbp) member bound to said second reagent and a second sbp member complementary to said first sbp member bound to said label wherein at least one of said first or second sbp members has only one specific binding site.
Claim 15
Dependent Claim 36 words
The method of claim 13 wherein said sbp member is an organic residue of molecular weight 125-1500 covalently bound to said second reagent and said complementary sbp member is a receptor specific for said organic residue.
Claim 16
Dependent Claim 20 words
The method of claim 13 wherein said label is selected from the group consisting of catalysts, fluorescers, chemiluminescers, and particles.
Claim 17
Dependent Claim 38 words
The method of claim 13 wherein said sbp member and said complementary sbp member are respectively a ligand of molecular weight 125-1500 and a receptor for said ligand, wherein a single ligand is bound to said second reagent.
Claim 18
Dependent Claim 15 words
The method of claim 16 wherein said ligand is biotin and said receptor is avidin.
Claim 19
Dependent Claim 29 words
The method of claim 1 wherein said means for rendering said second reagent detectable includes a label selected from the group consisting of catalysts, dyes, fluorescers, chemiluminescers, and particles.
Claim 20
Dependent Claim 25 words
The method of claim 18 wherein said label is selected from the group consisting of metal sols, carbon particles, polymeric particles, liposomes, and dye aggregates.
Claim 21
Dependent Claim 12 words
The method of claim 18 wherein said label is a fluorescent particle.
Claim 22
Dependent Claim 11 words
The method of claim 18 wherein a second label is employed.
Claim 23
Dependent Claim 37 words
The method of claim 21 wherein said label is a first enzyme and said second label is a second enzyme wherein said enzymes are related in that the product of one is the substrate for the other.
Claim 24
Dependent Claim 18 words
The method of claim 21 wherein said label is a fluorescer and said second label is a quencher.
Claim 25
Dependent Claim 27 words
The method of claim 21 wherein said second label is covalently or noncovalently bound to a compound capable of causing the non-covalent polymerization of said first reagent.
Claim 26
Independent Claim 115 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample, (A) a polymerization probe capable of hybridizing with said analyte and capable of being non-covalently polymerized, and (B) a detection probe capable of hybridizing with said analyte in a region other than that with which said non-covalent polymerization probe hybridizes and capable of being detected, which detection probe becomes bound to the polymerized polymerization probe only when said analyte is present in said sample, (b) causing the polymerization of said polymerization probe, and (c) determining whether said detection probe is bound to said polymerized polymerization probe.
Claim 27
Independent Claim 115 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample, (A) a polymerization probe capable of hybridizing with said analyte and capable of being non-covalently polymerized, and (B) a detection probe capable of hybridizing with said analyte in a region other than that with which said non-covalent polymerization probe hybridizes and capable of being detected, which detection probe becomes bound to the polymerized polymerization probe only when said analyte is present in said sample, (b) causing the polymerization of said polymerization probe, and (c) determining whether said detection probe is bound to said polymerized polymerization probe.
Claim 28
Dependent Claim 23 words
The method of claim 25 wherein a compound capable of causing the non-covalent polymerization of said polymerization probe is combined in Step b.
Claim 29
Dependent Claim 11 words
The method of claim 25 wherein said polynucleotide analyte is DNA.
Claim 30
Dependent Claim 11 words
The method of claim 25 wherein said polynucleotide analyte is RNA.
Claim 31
Dependent Claim 29 words
The method of claim 25 wherein in Step c said polymerized polymerization probe is examined for the presence of said detection probe by determining the presence of aggregated particles.
Claim 32
Dependent Claim 25 words
The method of claim 25 wherein said polynucleotide analyte is selected from the group consisting of genomes of viruses, bacteria, and fungi and fragments thereof.
Claim 33
Dependent Claim 10 words
The method of claim 30 wherein the fungi are molds.
Claim 34
Dependent Claim 29 words
The method of claim 25 wherein said polymerization probe comprises an oligonucleotide, complementary to a portion of said analyte, bound to means for rendering said polymerization probe non-covalently polymerizable.
Claim 35
Dependent Claim 19 words
The method of claim 32 wherein said means includes a repeating oligonucleotide sequence covalently bound to said polymerization probe.
Claim 36
Dependent Claim 17 words
The method of claim 32 wherein said oligonucleotide is a homopolymer having at least eight nucleotide residues.
Claim 37
Dependent Claim 20 words
The method of claim 32 wherein said oligonucleotide is a heteropolymer containing at least eight nucleotides comprising two non-complementary nucleotides.
Claim 38
Dependent Claim 39 words
The method of claim 32 wherein said means includes a specific binding pair (sbp) member bound to said polymerization probe and wherein a complementary sbp member having at least two specific binding sites is combined with said assay medium.
Claim 39
Dependent Claim 28 words
The method of claim 36 wherein said sbp member and said complementary sbp member are selected from the group consisting of antigen-antibody, avidin-biotin, IgG-protein A, DNA-DNA and DNA-RNA.
Claim 40
Dependent Claim 22 words
The method of claim 25 wherein said detection probe comprises an oligonucleotide bound to a means for rendering said detection probe detectable.
Claim 41
Dependent Claim 32 words
The method of claim 38 wherein said means includes a label selected from the group consisting of catalysts, dyes, fluorescers, chemiluminescers, particles, metal sols, carbon particles, polymeric particles, liposomes, and dye aggregates.
Claim 42
Dependent Claim 12 words
The method of claim 39 wherein said label is a fluorescent particle.
Claim 43
Dependent Claim 33 words
The method of claim 39 wherein said label is selected from the group consisting of catalysts, dyes, enzymes, coenzymes, fluorescers, quenchers, chemiluminescers, particles, metal sols, carbon particles, polymeric particles, lyposomes, and dye aggregates.
Claim 44
Dependent Claim 11 words
The method of claim 39 wherein a second label is employed.
Claim 45
Dependent Claim 37 words
The method of claim 42 wherein said label is a first enzyme and said second label is a second enzyme wherein said enzymes are related in that the product of one is the substrate for the other.
Claim 46
Dependent Claim 18 words
The method of claim 42 wherein said label is a fluorescer and said second label is a qunecher.
Claim 47
Dependent Claim 30 words
The method of claim 42 wherein said second label is bound to, or is capable of binding to, a compound capable of causing the non-covalent polymerization of said polymerization probe.
Claim 48
Dependent Claim 53 words
The method of claim 39 wherein said means includes a first specific binding pair (sbp) member bound to said detection probe and a second sbp member complementary to said first sbp member bound to said label wherein at least one of said first or second sbp members has a single specific binding site.
Claim 49
Dependent Claim 34 words
The member of claim 46 wherein said sbp member is an organic residue of molecular weight 125-1500 covalently bound to said detection probe and said complementary sbp member is specific for said organic residue.
Claim 50
Dependent Claim 35 words
The method of claim 46 wherein said sbp member and said complementary sbp member are respectively a ligand of molecular weight 125-1500 and a receptor wherein a single ligand is bound to said detection probe.
Claim 51
Dependent Claim 15 words
The method of claim 48 wherein said ligand is biotin and said receptor is avidin.
Claim 52
Independent Claim 146 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and (A) a polymerization probe comprising (1) a polynucleotide, hybridizable with a region of said analyte, conjugated to (2) means for non-covalently polymerizing said polymerization probe, (B) a detection probe comprising (1) a polynucleotide, hybridizable with a region of said analyte other than the region recognized by said polymerization probe, conjugated to (2) means for rendering said detection probe detectable, and (C) a polymerization agent capable of causing the non-covalent polymerization of said polymerization probe, under conditions for polymerizing said polymerization probe wherein said detection probe becomes bound to said polymerized polymerization probe only when said analyte is present in said sample, and (b) determining whether said detection probe has become bound to said polymerized polymerization probe.
Claim 53
Independent Claim 146 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and (A) a polymerization probe comprising (1) a polynucleotide, hybridizable with a region of said analyte, conjugated to (2) means for non-covalently polymerizing said polymerization probe, (B) a detection probe comprising (1) a polynucleotide, hybridizable with a region of said analyte other than the region recognized by said polymerization probe, conjugated to (2) means for rendering said detection probe detectable, and (C) a polymerization agent capable of causing the non-covalent polymerization of said polymerization probe, under conditions for polymerizing said polymerization probe wherein said detection probe becomes bound to said polymerized polymerization probe only when said analyte is present in said sample, and (b) determining whether said detection probe has become bound to said polymerized polymerization probe.
Claim 54
Dependent Claim 11 words
The method of claim 50 wherein said polynucleotide analyte is DNA.
Claim 55
Dependent Claim 11 words
The method of claim 50 wherein said polynucleotide analyte is RNA.
Claim 56
Dependent Claim 29 words
The method of claim 50 wherein in Step b said polymerized polymerization probe is examined for the presence of said detection probe by determining the presence of aggregated particles.
Claim 57
Dependent Claim 25 words
The method of claim 50 wherein said polynucleotide analyte is selected from the group consisting of genomes of viruses, bacteria, and fungi, and fragments thereof.
Claim 58
Dependent Claim 10 words
The method of claim 54 wherein the fungi are molds.
Claim 59
Dependent Claim 22 words
The method of claim 50 wherein said means for non-covalently polymerizing said polymerization probe includes a repeating oligonucleotide sequence covalently bound thereto.
Claim 60
Dependent Claim 23 words
The method of claim 56 wherein said repeating oligonucleotide sequence is a heteropolymer containing at least eight nucleotides and comprising two non-complementary nucleotides.
Claim 61
Dependent Claim 40 words
The method of claim 50 wherein said means for rendering said polymeric detection probe detectable includes a label selected from the group consisting of catalysts, dyes, fluorescers, quenchers, chemiluminescers, particles, metal sols, carbon particles, polymeric particles, liposomes, and dye aggregates.
Claim 62
Dependent Claim 12 words
The method of claim 58 wherein said label is a fluorescent particle.
Claim 63
Dependent Claim 11 words
The method of claim 58 wherein a second label is employed.
Claim 64
Dependent Claim 37 words
The method of claim 60 wherein said label is a first enzyme and said second label is a second enzyme wherein said enzymes are related in that the product of one is the substrate for the other.
Claim 65
Dependent Claim 18 words
The method of claim 60 wherein said label is a fluorescer and said second label is a quencher.
Claim 66
Dependent Claim 30 words
The method of claim 60 wherein said second label is bound to, or may become bound to, a polymerization agent capable of causing the non-covalent polymerization of said polymerization probe.
Claim 67
Dependent Claim 60 words
The method of claim 58 wherein said means for rendering said polymeric detection probe detectable includes a first specific binding pair (sbp) member bound to said detection probe and a second sbp member complementary to said first sbp member bound to said label wherein at least one of said first or second sbp members has a single specific binding site.
Claim 68
Dependent Claim 28 words
The method of claim 64 wherein said sbp member and said complementary sbp member are selected from the group consisting of antigen-antibody, avidin-biotin, IgG-protein A, DNA-DNA, and DNA-RNA.
Claim 69
Dependent Claim 30 words
The method of claim 64 wherein said label is selected from the group consisting of catalysts, dyes, fluorescers, chemiluminescers, particles, metal sols, carbon particles, polymeric particles, liposomes, and dye aggregates.
Claim 70
Independent Claim 192 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and (A) a polymerization probe comprising (1) a polynucleotide, hybridizable with a region of said analyte, bound to (2) a repeating oligonucleotide sequence selected from the group consisting of a homopolymer having at least eight nucleotide residues and a heteropolymer consisting of two non-complementary nucleotides in a nucleotide sequence at least eight nucleotides in length, (B) a detection probe comprising (1) a polynucleotide, hybridizable with a region of said analyte other than the region recognized by said polymerization probe, bound to (2) a label selected from the group consisting of catalysts, enzymes, coenzymes, dyes, fluorescers, chemiluminescers, and particles, and (C) a polymerizing agent comprising a repeating oligonucleotide sequence complementary to said repeating oligonucleotide sequence bound to said polymerization probe, (b) causing polymerization of said polymerization probe under conditions wherein said detection probe becomes bound to said polymerized polymerization probe only if said analyte is present in said sample, and (c) determining whether said detection probe has become bound to said polymerized polymerization probe.
Claim 71
Independent Claim 192 words
A method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which method comprises (a) combining in an assay medium said sample and (A) a polymerization probe comprising (1) a polynucleotide, hybridizable with a region of said analyte, bound to (2) a repeating oligonucleotide sequence selected from the group consisting of a homopolymer having at least eight nucleotide residues and a heteropolymer consisting of two non-complementary nucleotides in a nucleotide sequence at least eight nucleotides in length, (B) a detection probe comprising (1) a polynucleotide, hybridizable with a region of said analyte other than the region recognized by said polymerization probe, bound to (2) a label selected from the group consisting of catalysts, enzymes, coenzymes, dyes, fluorescers, chemiluminescers, and particles, and (C) a polymerizing agent comprising a repeating oligonucleotide sequence complementary to said repeating oligonucleotide sequence bound to said polymerization probe, (b) causing polymerization of said polymerization probe under conditions wherein said detection probe becomes bound to said polymerized polymerization probe only if said analyte is present in said sample, and (c) determining whether said detection probe has become bound to said polymerized polymerization probe.
Claim 72
Dependent Claim 26 words
The method of claim 67 wherein said polymerized polymerization probe is examined for the presence of said detection probe by determining the presence of aggregated particles.
Claim 73
Dependent Claim 11 words
The method of claim 67 wherein a second label is employed.
Claim 74
Dependent Claim 37 words
The method of claim 69 wherein said label is a first enzyme and said second label is a second enzyme wherein said enzymes are related in that the product of one is the substrate for the other.
Claim 75
Dependent Claim 18 words
The method of claim 69 wherein said label is a fluorescer and said second label is a quencher.
Claim 76
Dependent Claim 20 words
The method of claim 69 wherein said second label is conjugated to, or may become conjugated to, said polymerization agent.
Claim 77
Dependent Claim 63 words
In an assay method for the detection of a polynucleotide analyte in a sample suspected of containing said analyte, said method comprising the steps of combining said sample with a reagent capable of hybridizing with said analyte and determining the extent of such hybridization, the improvement which comprises employing a nucleotide sequence to cause non-covalent polymerization to determine the extent of such hybridization.
Claim 78
Dependent Claim 63 words
In an assay method for the detection of a polynucleotide analyte in a sample suspected of containing said analyte, said method comprising the steps of combining said sample with a reagent capable of hybridizing with said analyte and determining the extent of such hybridization, the improvement which comprises employing a nucleotide sequence to cause non-covalent polymerization to determine the extent of such hybridization.
Claim 79
Dependent Claim 11 words
The method of claim 73 wherein said polynucleotide analyte is DNA.
Claim 80
Dependent Claim 11 words
The method of claim 73 wherein said polynucleotide analyte is RNA.
Claim 81
Dependent Claim 15 words
The method of claim 73 wherein said polynucleotide analyte is the genome of herpes virus.
Claim 82
Dependent Claim 18 words
The method of claim 73 wherein said agglutination employs repeating oligonucleotide sequences having at least eight nucleotide residues.
Claim 83
Dependent Claim 23 words
The method of claim 73 wherein said nucleotide sequence is a heteropolymer at least eight nucleotides in length comprising two repeating non-complementary nucleotides.
Claim 84
Dependent Claim 25 words
The method of claim 73 wherein said polynucleotide analyte is selected from the group consisting of genomes of viruses, bacteria, and fungi, and fragments thereof.
Claim 85
Dependent Claim 10 words
The method of claim 79 wherein the fungi are molds.
Claim 86
Dependent Claim 119 words
A kit useful in a method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which kit comprises in packaged combination (a) polymerization probe comprising (1) a polynucleotide, complementary with a region of said analyte to an extent sufficient to hybridize to said analyte, conjugated to (2) means for rendering said polymerization probe non-covalently polymerizable, (b) a detection probe comprising (1) a polynucleotide, complementary with a region of said analyte, to an extent sufficient to hybridize to said analyte, other than the region recognized by said polymerization probe, conjugated to (2) means for rendering said detection probe detectable, and (c) a polymerization agent for causing the non-covalent polymerization of said polymerization probe.
Claim 87
Dependent Claim 119 words
A kit useful in a method for determining the presence of a polynucleotide analyte in a sample suspected of containing said analyte, which kit comprises in packaged combination (a) polymerization probe comprising (1) a polynucleotide, complementary with a region of said analyte to an extent sufficient to hybridize to said analyte, conjugated to (2) means for rendering said polymerization probe non-covalently polymerizable, (b) a detection probe comprising (1) a polynucleotide, complementary with a region of said analyte, to an extent sufficient to hybridize to said analyte, other than the region recognized by said polymerization probe, conjugated to (2) means for rendering said detection probe detectable, and (c) a polymerization agent for causing the non-covalent polymerization of said polymerization probe.
Claim 88
Dependent Claim 33 words
The kit of claim 81 wherein the means for rendering said polymerization probe non-covalently polymerizable includes a repeating oligonucleotide sequence and said polymerization agent is a repeating oligonucleotide sequence complementary to said sequence.
Claim 89
Dependent Claim 41 words
The kit of claim 81 wherein said means for rendering said polymerization probe non-covalently polymerizable includes a specific binding pair (sbp) member bound thereto and wherein said kit further comprises a complementary sbp member having at least two specific binding sites.
Claim 90
Dependent Claim 28 words
The kit of claim 83 wherein said sbp member and said complementary sbp member are selected from the group consisting of antigen-antibody, avidin-biotin, IgG-protein A, DNA-DNA, and DNA-RNA.
Claim 91
Dependent Claim 20 words
The kit of claim 81 wherein said means for rendering said polymerization probe non-covalently polymerizable includes a repeating oligonucleotide sequence.
Claim 92
Dependent Claim 19 words
The kit of claim 85 wherein said repeating nucleotide sequence is a homopolymer having at least eight nucleotide residues.
Claim 93
Dependent Claim 23 words
The kit of claim 85 wherein said repeating nucleotide sequence is a heteropolymer having at least eight nucleotide residues comprising two non-complementary nucleotides.
Claim 94
Dependent Claim 33 words
The kit of claim 81 wherein said means for rendering said polymeric detection probe detectable includes a label selected from the group consisting of catalysts, enzymes, coenzymes, dyes, fluorescers, quenchers, chemiluminescers and particles.
Claim 95
Dependent Claim 25 words
The kit of claim 88 which further includes a second label conjugated to a compound capable of causing the non-covalent polymerization of said polymerization probe.
Claim 96
Dependent Claim 37 words
The kit of claim 89 wherein said label is a first enzyme and said second label is a second enzyme wherein said enzymes are related in that the product of one is the substrate for the other.
Claim 97
Dependent Claim 18 words
The kit of claim 89 wherein said label is a fluorescer and said second label is a quencher.
Claim 98
Dependent Claim 63 words
The kit of claim 88 wherein said means for rendering said detection probe detectable includes a first specific binding pair (sbp) member bound to said detection probe and said kit further comprises a second sbp member complementary to said first sbp member bound to said label wherein at least one of said first and second sbp members has a single specific binding site.
Claim 99
Dependent Claim 35 words
The kit of claim 92 wherein said sbp member is an organic residue of molecular weight 125-1500 covalently bound to said detection probe and said complementary sbp member is a receptor specific for said residue.
Claim 100
Dependent Claim 24 words
The kit of claim 92 wherein said label is selected from the group consisting of catalyts, enzymes, coenzymes, dyes, fluorescers, quenchers, chemiluminescers, and particles.
Claim 101
Dependent Claim 35 words
The kit of claim 92 wherein said sbp member and said complementary sbp members are respectively a ligand of molecular weight 125-1500 and a receptor wherein a single ligand is bound to the detection probe.
Claim 102
Dependent Claim 16 words
The kit of claim 95 wherein said organic residue is biotin and said receptor is avidin.

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