EP0163220A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Nucleic acid hybridization assay employing detectable anti-hybrid antibodies
Assignee: Bayer Corp
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Filing: 1985-05-18
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Granted: —
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Expires: 2005-05-18
Abstract
Abstract A nucleic acid hybridization assay employing an immobilized or immobilizable polynucleotide probe selected to form DNA'RNA or RNA'RNA hybrids with the particular polynucleotide sequence to be determined. Resulting hybrids are detected by binding of an antibody reagent, preferably labeled with a detectable chemical group, selective for binding the hybrids in the presence of the single stranded sample and probe nucleic acids. No immobilization or labeling of sample nucleic acids is necessary and hybridization can be performed entirely in solution.
Inventors
- 👤 Robert J. Carrico
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-03 14:37
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6804; C12Q1/6876; C12Q1/6888; C12Q1/689; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/5308; G01N33/543; G01N33/54306
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
159 words
A method for determining a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, comprising the steps of combining the test medium with an immobilized or immobilizable polynucleotide probe comprising at least one single stranded base sequence which is substantially complementary to the sequence to be determined under conditions favorable to hybridization between the sequence to be determined and the complementary probe sequence, immobilizing the probe where provided in an immobilizable form, and detecting hybridized probe, characterized in that the complementary probe sequence is (i) substantially composed of RNA when the sequence to be determined is RNA or DNA, or (ii) substantially composed of DNA or RNA when the sequence to be determined is RNA, and hybridized probe is determined by adding an antibody reagent capable of binding to DNA'RNA or RNA'RNA duplexes formed between the sequence to be determined and the complementary probe sequence and determining the antibody reagent that becomes bound to such duplexes.
Claim 2
Independent Claim
159 words
A method for determining a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, comprising the steps of combining the test medium with an immobilized or immobilizable polynucleotide probe comprising at least one single stranded base sequence which is substantially complementary to the sequence to be determined under conditions favorable to hybridization between the sequence to be determined and the complementary probe sequence, immobilizing the probe where provided in an immobilizable form, and detecting hybridized probe, characterized in that the complementary probe sequence is (i) substantially composed of RNA when the sequence to be determined is RNA or DNA, or (ii) substantially composed of DNA or RNA when the sequence to be determined is RNA, and hybridized probe is determined by adding an antibody reagent capable of binding to DNA'RNA or RNA'RNA duplexes formed between the sequence to be determined and the complementary probe sequence and determining the antibody reagent that becomes bound to such duplexes.
Claim 3
Dependent Claim
19 words
The method of Claim 1 wherein the probe is in an immobilized form when contacted with the test medium.
Claim 4
Dependent Claim
30 words
The method of Claim 1 wherein the probe comprises a specific binding site and is immobilizable by contact with an immobilized form of a binding partner for such binding site.
Claim 5
Dependent Claim
41 words
The method of Claim 1 wherein the probe is in a soluble form when contacted with the test medium and is immobilizable when hybridized to the sequence to be determined by contact with an adsorbent which adsorbs double stranded nucleic acids.
Claim 6
Independent Claim
38 words
The method of any one of Claims 1-4 wherein the antibody reagent is labeled with a detectable chemical group selected from an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand, or a radioisotope.
Claim 7
Dependent Claim
27 words
The method of Claim 1 wherein the test medium comprises a biological sample which has been subjected to conditions to release and denature nucleic acids present therein.
Claim 8
Dependent Claim
111 words
A reagent system for detecting a particular polynucleotide sequence in a test medium by nucleic acid hybridization, comprising an immobilized or immobilizable polynucleotide probe comprising at least one single stranded base sequence which is substantially complementary to the sequence to be determined, characterized in that the complementary probe sequence is (i) substantially composed of RNA when the sequence to be determined is RNA or DNA, or (ii) is substantially composed of DNA or RNA when the sequence to be determined is RNA; and the reagent system additionally comprises an antibody reagent capable of binding to DNA'RNA or RNA'RNA duplexes formed between the sequence to be determined and the complementary probe sequence.
Claim 9
Dependent Claim
111 words
A reagent system for detecting a particular polynucleotide sequence in a test medium by nucleic acid hybridization, comprising an immobilized or immobilizable polynucleotide probe comprising at least one single stranded base sequence which is substantially complementary to the sequence to be determined, characterized in that the complementary probe sequence is (i) substantially composed of RNA when the sequence to be determined is RNA or DNA, or (ii) is substantially composed of DNA or RNA when the sequence to be determined is RNA; and the reagent system additionally comprises an antibody reagent capable of binding to DNA'RNA or RNA'RNA duplexes formed between the sequence to be determined and the complementary probe sequence.
Claim 10
Dependent Claim
18 words
The reagent system of Claim 7 wherein the probe is immobilized by being fixed to a solid support.
Claim 11
Dependent Claim
38 words
The reagent system of Claim 7 wherein the probe is immobilizable and comprises a specific binding site, and wherein the reagent system additionally comprises an immobilized form of a binding partner for the binding site on the probe.
Claim 12
Dependent Claim
39 words
The reagent system of any one of Claims 8-10 wherein the antibody reagent is labeled with a detectable chemical group selected from an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand, or a radioisotope.
References Cited (Backward Citations)
No citation records indexed.
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