Method for detecting a target molecule in a sample using a nucleic acid ligand
First Claim
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1. A method for detecting the presence or absence of a target molecule in a sample which may contain said target molecule comprising:
- a) exposing said sample which may contain said target molecule to a non-naturally occurring nucleic acid ligand of said target molecule, wherein said nucleic acid ligand is not a nucleic acid having the known physiological function of being bound by said target molecule, under conditions wherein a target molecule;
nucleic acid ligand binding pair is formed if said target molecule is present, wherein said binding is not due to Watson/Crick base pairing; and
b) determining whether said target molecule;
nucleic acid ligand binding pair is formed, thereby detecting the presence or absence of the target molecule in the sample.
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Abstract
This invention discloses a method of detecting the presence or absence of a target molecule in a sample and a method of measuring the amount of a target molecule in a sample using nucleic acid ligands. In a preferred embodiment the nucleic acid ligands are identified by the method of the invention referred to as the Systematic Evolution of Ligands by EXponential Enrichment (SELEX), wherein a candidate mixture of nucleic acids are iteratively enriched in high affinity nucleic acids and amplified for further partitioning.
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Citations
57 Claims
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1. A method for detecting the presence or absence of a target molecule in a sample which may contain said target molecule comprising:
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a) exposing said sample which may contain said target molecule to a non-naturally occurring nucleic acid ligand of said target molecule, wherein said nucleic acid ligand is not a nucleic acid having the known physiological function of being bound by said target molecule, under conditions wherein a target molecule;
nucleic acid ligand binding pair is formed if said target molecule is present, wherein said binding is not due to Watson/Crick base pairing; andb) determining whether said target molecule;
nucleic acid ligand binding pair is formed, thereby detecting the presence or absence of the target molecule in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for detecting the presence or absence of a target molecule in a sample which may contain said target molecule comprising:
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a) identifying a nucleic acid ligand from a candidate mixture of nucleic acids, said nucleic acid ligand being a ligand of said target molecule, by a method comprising; i) contacting the candidate mixture with said target molecule, wherein nucleic acids having an increased affinity to said target molecule relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; ii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and iii) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids, whereby an increased affinity nucleic acid ligand may be identified; b) exposing said sample which may contain said target molecule to said increased affinity nucleic acid ligand of said target molecule under conditions wherein a target molecule;
nucleic acid ligand binding pair is formed if said target molecule is present; andc) determining whether said target molecule;
nucleic acid ligand binding pair is formed, thereby detecting the presence or absence of the target molecule in the sample. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. A method for measuring the amount of a target molecule in a sample which may contain said target molecule comprising:
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a) exposing said sample which may contain said target molecule to a nucleic acid ligand of said target molecule, wherein said nucleic acid ligand is not a nucleic acid having the known physiological function of being bound by the target molecule, under conditions wherein a target molecule;
nucleic acid ligand binding pair is formed;b) determining whether said target molecule;
nucleic acid ligand binding pair is formed wherein said binding is not due to Watson/Crick base pairing; andc) measuring the amount of the target molecule in the sample by quantitatively detecting the nucleic acid ligand. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42)
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43. A method for measuring the amount of a target molecule in a sample which may contain said target molecule comprising:
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a) identifying a nucleic acid ligand from a candidate mixture of nucleic acids, said nucleic acid ligand being a ligand of said target molecule, by a method comprising; i) contacting the candidate mixture with said target molecule, wherein nucleic acids having an increased affinity to said target molecule relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; ii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and iii) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids, whereby an increased affinity nucleic acid ligand may be identified; b) exposing said sample which may contain said target molecule to said increased affinity nucleic acid ligand of said target molecule under conditions wherein a target molecule;
nucleic acid ligand binding pair is formed;c) determining whether said target molecule;
nucleic acid ligand binding pair is formed; andd measuring the amount of the target molecule in the sample, by quantitatively detecting the nucleic acid ligand. - View Dependent Claims (44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57)
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Specification