Process for detecting a nucleic acid of interest using an oligo- or polynucleotide probe and intercalators
First Claim
1. A process for detecting the presence of a nucleic acid sequence of interest comprising:
- (a) providing;
(i) an oligo- or polynucleotide which is specifically hybridizable with said nucleic acid sequence of interest, said oligo or polynucleotide comprising at least one lanthanide metal or at least one fluorophore, each acting as either an energy donor or an energy acceptor;
(ii) a solid matrix having at least one surface to which is fixed or immobilized a first intercalator capable of capturing double-stranded nucleic acid, and (iii) a second intercalator, which may or may not comprise at least one fluorophore, said second intercalator or said fluorophore, each acting as either an energy donor or an energy acceptor;
(b) forming a complex comprising;
said oligo- or polynucleotide (i), said solid matrix (ii), said second intercalator (iii), and said nucleic acid sequence of interest if contained in said sample; and
(c) detecting the presence of said nucleic acid sequence of interest by detecting any energy emitted from said energy acceptor.
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Accused Products
Abstract
Disclosed is a nucleic acid hybridization assay composition for detecting the presence or absence of a target oligo- or polynucleotide in a sample. The composition comprises: a solid matrix having at least one surface which is substituted with a first intercalator capable of binding dsDNA, dsRNA, or DNA-RNA hybrids; a second intercalator, which may or may not comprise at least one fluorophore, said intercalator or said fluorophore, each acting as either an energy donor or an energy acceptor: and an oligo- or polynucleotide probe which is specifically hybridizable with the target oligo- or polynucleotide and has directly or indirectly bound thereto, at least one lanthanide metal chelate or at least one fluorophore, each acting as either an energy donor or an energy acceptor. Also disclosed are a method and kit for its use.
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Citations
24 Claims
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1. A process for detecting the presence of a nucleic acid sequence of interest comprising:
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(a) providing;
(i) an oligo- or polynucleotide which is specifically hybridizable with said nucleic acid sequence of interest, said oligo or polynucleotide comprising at least one lanthanide metal or at least one fluorophore, each acting as either an energy donor or an energy acceptor;
(ii) a solid matrix having at least one surface to which is fixed or immobilized a first intercalator capable of capturing double-stranded nucleic acid, and (iii) a second intercalator, which may or may not comprise at least one fluorophore, said second intercalator or said fluorophore, each acting as either an energy donor or an energy acceptor;
(b) forming a complex comprising;
said oligo- or polynucleotide (i), said solid matrix (ii), said second intercalator (iii), and said nucleic acid sequence of interest if contained in said sample; and
(c) detecting the presence of said nucleic acid sequence of interest by detecting any energy emitted from said energy acceptor. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A process for detecting the presence of a nucleic acid sequence of interest in a sample comprising:
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(a) providing a solid matrix having at least one surface to which is attached a first intercalator capable of capturing double-stranded nucleic acid;
(b) contacting under hybridizable conditions said solid matrix with the sample suspected of containing said nucleic acid sequence of interest and an oligo- or polynucleotide which is specifically hybridizable therewith, said oligo or polynucleotide comprising at least one lanthanide metal or at least one fluorophore, each acting as either an energy donor or an energy acceptor, to form a complex which is captured by said solid matrix;
(c) contacting said captured complex with a second intercalator, which may or may not comprise at least one fluorophore, said second intercalator or said fluorophore, each acting as either an energy donor or an energy acceptor, and (d) detecting the presence of said nucleic acid sequence of interest by detecting any energy emitted from said energy acceptor. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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17. A process for detecting the presence of a nucleic acid sequence of interest comprising:
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(a) contacting a sample suspected of containing said nucleic acid sequence of interest with an oligo- or polynucleotide hybridizable therewith to form a hybrid, said oligo- or polynucleotide having directly or indirectly bound thereto a lanthanide metal or a fluorophore, each independently acting as either an energy donor or an energy acceptor;
(b) contacting said hybrid with a solid matrix having at least one surface to which is attached a first intercalator capable of capturing double-stranded nucleic acid;
(c) permitting capture of said hybrid to form a complex;
(d) contacting said complex with a second intercalator, which may or may not comprise at least one fluorophore, said second intercalator or said fluorophore, each acting as either an energy donor or an energy acceptor; and
(e) detecting the presence of said nucleic acid sequence of interest by detecting any energy emitted from said energy acceptor. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24)
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Specification