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EP0147665A1
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Nucleic acid probe, test method and reagent system for detecting a polynucleotide sequence and antibody therefor
Assignee: Molecular Diagnostics Inc
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Filing: 1984-11-30
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Granted: —
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Expires: 2004-11-30
Abstract
Abstract A nucleic acid detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion preferably including a recognition site for binding by a particular protein. Such recognition site can be a region of singly or doubly stranded nucleic acid specific for a particular nucleic acid binding protein such as lac repressor protein or can be a modified nucleic acid region such as a unique antigenic determinant introduced by interaction of the region with a modifier compound such as an intercalating agent or a platinium-containing ligand. The probe-binding protein can be labeled for ease of detection and in the case of an antigenic determinant binding site can be labeled antibody.
Inventors
- 👤 Nanibhushan Dattagupta
- 👤 Peter M. M. Rae
- 👤 William J. Knowles
- 👤 Donald M. Crothers
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 19:00
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6876; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/811
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
38 words
A nucleic acid detection probe comprising a hybridizable single stranded protion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein.
Claim 2
Dependent Claim
38 words
A nucleic acid detection probe comprising a hybridizable single stranded protion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein.
Claim 3
Dependent Claim
24 words
A detection probe according to claim 1, characterized in that the hybridizable portion is continuous with one of the strands of the non-hybridizable portion.
Claim 4
Dependent Claim
40 words
A detection probe according to claim 1 or 2, characterized in that the non-hybridizable portion includes a recognition site for a protein selected from the group of lactose repressor, galactose repressor, lambda repressor, catabolite gene activator protein and Cro protein.
Claim 5
Dependent Claim
57 words
A detection probe according to any of claims 1 to 3, characterized in that the non-hybridizable portion has been modified to create the protein recognition site by contact or reaction with a modifier compound which introduces a unique antigenic determinant into the non-hybridizable portion, the modifier compound preferably being an intercalating agent, a platinium-containing ligand or salt.
Claim 6
Dependent Claim
53 words
A detection probe according to any of claims 1 to 4, characterized in that it includes the particular protein bound to the protein-specific site of the non-hybridizable portion, the protein preferably carrying a label selected from an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand, or a radioisotope.
Claim 7
Dependent Claim
19 words
A detection probe according to claim 5, characterized in that the protein is an antibody, or a fragment thereof.
Claim 8
Independent Claim
129 words
A method for detecting a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, characterized in that it comprises the steps of: a) combining the test medium with a nucleic acid detection probe comprising at least one hybridizable single stranded base sequence which is substantially complementary to the sequence to be detected and a non-hybridizable double-stranded portion having a recognition site for binding by a particular protein, under conditions favorable to hybridization between the sequence to be detected and the complementary hybridizable sequence in the probe, b) separating hybridized probe from unhybridized probe, and c) adding to separated hybridized probe, the particular protein which binds the recognition site on the non-hybridizable portion of the probe and determining the protein which beeches bound to the solid support.
Claim 9
Independent Claim
129 words
A method for detecting a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, characterized in that it comprises the steps of: a) combining the test medium with a nucleic acid detection probe comprising at least one hybridizable single stranded base sequence which is substantially complementary to the sequence to be detected and a non-hybridizable double-stranded portion having a recognition site for binding by a particular protein, under conditions favorable to hybridization between the sequence to be detected and the complementary hybridizable sequence in the probe, b) separating hybridized probe from unhybridized probe, and c) adding to separated hybridized probe, the particular protein which binds the recognition site on the non-hybridizable portion of the probe and determining the protein which beeches bound to the solid support.
Claim 10
Dependent Claim
111 words
A method according to claim 7 which is a solid phase hybridization technique, wherein the single stranded nucleic acids from the test medium are immobilized on a solid support and wherein the protein associated with the solid support is determined or which is a solid phase sandwich hybridization technique, wherein the test medium is combined with first and second nucleic acid probes each comprising at least one hybridizable single stranded base sequence which is substantially complementary to a mutually exclusive portion of the sequence to be detected and wherein one of the probes is immobilized on a solid support and the other has said non-hybridizable portion having said protein recognition site.
Claim 11
Dependent Claim
50 words
A reagent system for detecting a particular polynucleotide sequence in a test medium, comprising: a) a nucleic acid detection probe according to any of claims 1 to 4, and b) said particular protein capable of binding to said recognition site on the detection probe, the protein preferably carrying a label.
Claim 12
Dependent Claim
38 words
A reagent system according to claim 9, characterized in that the protein is an antibody, or a fragment thereof, labeled with an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand, or a radio-isotope.
Claim 13
Dependent Claim
33 words
An antibody, or fragment thereof, capable of binding with an intercalation complex formed between double stranded nucleic acid and a nucleic acid, and preferably substantially incapable of binding to single stranded nucleic acids.
Claim 14
Dependent Claim
33 words
An antibody, or fragment thereof, capable of binding with an intercalation complex formed between double stranded nucleic acid and a nucleic acid, and preferably substantially incapable of binding to single stranded nucleic acids.
Claim 15
Dependent Claim
38 words
An antibody or fragment thereof according to claim 11, characterizedin that it is labeled with a detectable chemical group such as an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand, or a radioisotope.
References Cited (Backward Citations)
EP0079139A1
Cited
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