Patent EP0232967A2 successfully refreshed from Google Patents.
EP0232967A2
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EP
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Competitive homogeneous Assay
Assignee: BP Corp North America Inc
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Filing: 1987-01-09
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Granted: —
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Expires: 2007-01-09
Abstract
Abstract Methods and compositions for performing assays for target polynucleotide strands include contacting a sample with a reagent which includes a first and a second polynucleotide probe. The first and second probes are capable of assuming a first position wherein the probes are bound to each other and a second position wherein the probes are bound to a target. The probes include label moieties capable of interacting to produce a signal indicative of the probes being in one of the two positions.
Inventors
- 👤 Larry Edward Morrison
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 21:31
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818; C07; C07H; C07H21/00; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
161 words
A method for assaying a sample for target polynucleotides comprising: (a) contacting sample with reagent under binding conditions wherein said reagent includes a first polynucleotide probe and a second polynucleotide probe, said first and second probes capable of assuming a first position wherein said first and second probes are bound to each other and at least one of said probes capable of assuming a second position wherein said probe strand is bound to said target, said first probe and second probe including a first label moiety associated with one of said probes and including a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said first and second probes are bound to each other to produce a signal capable of detection characteristic of the probe in one of said two positions. (b) monitoring said sample for said signal, the presence of which is related to the presence of target in said sample.
Claim 2
Independent Claim
161 words
A method for assaying a sample for target polynucleotides comprising: (a) contacting sample with reagent under binding conditions wherein said reagent includes a first polynucleotide probe and a second polynucleotide probe, said first and second probes capable of assuming a first position wherein said first and second probes are bound to each other and at least one of said probes capable of assuming a second position wherein said probe strand is bound to said target, said first probe and second probe including a first label moiety associated with one of said probes and including a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said first and second probes are bound to each other to produce a signal capable of detection characteristic of the probe in one of said two positions. (b) monitoring said sample for said signal, the presence of which is related to the presence of target in said sample.
Claim 3
Independent Claim
35 words
The method of Claim l wherein said first label moiety is located at the 3′-terminus of one of said probes and said second label moiety is located at the 5′-terminus of the said opposite probe.
Claim 4
Dependent Claim
17 words
The method of Claim 1 or Claim 2 wherein each probe has a plurality of label moieties.
Claim 5
Independent Claim
19 words
The method of any preceding claim wherein each such label moiety is located at the termini of said probes.
Claim 6
Independent Claim
24 words
The method of any preceding claim wherein said first label moiety is associated to said probe by an aminoalkyl derivative of a nucleic acid.
Claim 7
Dependent Claim
14 words
The method of Claim 5 wherein said derivative includes an aminoalkyl derivative of adenine.
Claim 8
Dependent Claim
14 words
The method of Claim 6 wherein said derivative includes an aminohexyl derivative of adenine.
Claim 9
Dependent Claim
10 words
The method of Claim 6 wherein said derivative includes 8-(6-aminohexyl)-aminoadenosine-5′-monophosphate.
Claim 10
Independent Claim
25 words
The method of any of Claims 1 to 7 wherein said label moiety at the 3′ terminus is a fluorescent derivative of a nucleic acid.
Claim 11
Dependent Claim
14 words
The method of Claim 9 wherein the label moiety includes a derivative of l-N⁶-ethenoadenosine-5′-monophosphate.
Claim 12
Independent Claim
50 words
A method for associating an amine-reactive moiety to the 3′-terminus of a polynucleotide comprising: (a) reacting said polynucleotide with an aminoalkyl derivative of a nucleic acid in the presence of the enzyme terminal transferase under reacting conditions. (b) reacting the amino group of the aminoalkyl derivative with said amine-reactive moiety.
Claim 13
Independent Claim
50 words
A method for associating an amine-reactive moiety to the 3′-terminus of a polynucleotide comprising: (a) reacting said polynucleotide with an aminoalkyl derivative of a nucleic acid in the presence of the enzyme terminal transferase under reacting conditions. (b) reacting the amino group of the aminoalkyl derivative with said amine-reactive moiety.
Claim 14
Dependent Claim
15 words
The method of Claim 11 wherein said aminoalkyl derivative includes an aminoalkyl derivative of adenine.
Claim 15
Dependent Claim
15 words
The method of Claim 11 or Claim 12 wherein said aminoalkyl derivative is a ribonucleotide.
Claim 16
Independent Claim
15 words
The method of any of Claims 11 to 13 wherein said aminoalkyl derivative includes 8-(6-aminohexyl)-aminoadenoine-5′-triphosphate.
Claim 17
Independent Claim
90 words
A method for preparing polynucleotide probes including a first probe and second probe, said first and second probes capable of assuming a first position wherein said first probe is bound to said second probe and a second position wherein at least one of said probes is bound to a target, comprising: splicing polynucleotide segments having base sequences substantially identical to said target sequences into amplification means to form multiple copies of said polynucleotide segments, isolating said segments, and associating label moieties to the termini of the segments to form probes.
Claim 18
Independent Claim
90 words
A method for preparing polynucleotide probes including a first probe and second probe, said first and second probes capable of assuming a first position wherein said first probe is bound to said second probe and a second position wherein at least one of said probes is bound to a target, comprising: splicing polynucleotide segments having base sequences substantially identical to said target sequences into amplification means to form multiple copies of said polynucleotide segments, isolating said segments, and associating label moieties to the termini of the segments to form probes.
Claim 19
Dependent Claim
13 words
The method of Claim 15 wherein amplication means include plasmids and phage particles.
Claim 20
Dependent Claim
34 words
The method of Claim 15 or Claim 16 wherein said segments, after isolation, are subjected to restriction digestion to form further subsegments, and said subsections are associated with said label moieties to form probes.
Claim 21
Independent Claim
49 words
The method of any of Claims 15 to 17 wherein a said first label moiety is associated with the 3′-termini by reacting said segments with an aminoalkyl derivative of a nucleotide in the presence of terminal transferase under reacting conditions, and reacting said aminoalkyl group with a label moiety.
Claim 22
Dependent Claim
45 words
The method of Claim 18 wherein a second label moiety is associated with the 5′-termini by reacting said segments with a bifunctional alkyl amine to form a segment with an amino alkyl group and thereafter reacting a second label moiety with said amino alkyl group.
Claim 23
Dependent Claim
135 words
A kit for assaying a sample for target polynucleotides comprising reagent, wherein said reagent includes a first polynucleotide probe and a second polynucleotide probe, said first and second probes capable of assuming a first position wherein said first and second probe are bound to each other and at least one of said probes capable of assuming a second position wherein said probe strand is bound to said target, said first probe and second probe including a first label moiety associated with one of said probes and including a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said first and second probes are bound to each other to produce a signal capable of detection which is characteristic of the probe in one of said two positions.
Claim 24
Dependent Claim
135 words
A kit for assaying a sample for target polynucleotides comprising reagent, wherein said reagent includes a first polynucleotide probe and a second polynucleotide probe, said first and second probes capable of assuming a first position wherein said first and second probe are bound to each other and at least one of said probes capable of assuming a second position wherein said probe strand is bound to said target, said first probe and second probe including a first label moiety associated with one of said probes and including a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said first and second probes are bound to each other to produce a signal capable of detection which is characteristic of the probe in one of said two positions.
Claim 25
Dependent Claim
25 words
The kit of Claim 2l wherein at least one or said probes and/or label moieties is as defined in any of Claims 2 to 9.
Claim 26
Independent Claim
139 words
An apparatus for performing homogeneous competitive assays for polynucleotide targets in a sample, chamber means adapted for receiving reagent and sample, wherein said reagent includes a first probe and a second probe which are capable of assuming a first position wherein the first probe is bound to said second probe; a second position wherein at least one of said probes is bound to said target, said probes having at least one label moiety associated with one probe and a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said probes are in said first position to produce a signal upon excitation of one of said label moieties which signal is indicative of one of said positions; means for exciting one of said label moieties; and means for detecting said signal.
Claim 27
Independent Claim
139 words
An apparatus for performing homogeneous competitive assays for polynucleotide targets in a sample, chamber means adapted for receiving reagent and sample, wherein said reagent includes a first probe and a second probe which are capable of assuming a first position wherein the first probe is bound to said second probe; a second position wherein at least one of said probes is bound to said target, said probes having at least one label moiety associated with one probe and a second label moiety associated with said opposite probe, said first and second label moieties capable of interacting when said probes are in said first position to produce a signal upon excitation of one of said label moieties which signal is indicative of one of said positions; means for exciting one of said label moieties; and means for detecting said signal.
Claim 28
Dependent Claim
25 words
The apparatus of Claim 22 wherein said label moieties are fluorophores and said means for exciting one of said label moieties include a light source.
Claim 29
Dependent Claim
36 words
The apparatus of Claim 22 wherein at least one of said label moieties is a chemiluminescent agent and said means for exciting one of said label moieties include means for introducing chemiluminescent cofactors into said chamber.
Claim 30
Independent Claim
17 words
The apparatus of any of Claims 22 to 24 wherein said detection means include a light detector.
References Cited (Backward Citations)
No citation records indexed.
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