Nucleic acid assay and method of detecting the presence of a nucleic acid sequence
OwnerThor W. Nilseninventor
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Abstract
The present invention is directed to an assay and a method of detecting the presence of a specific sample nucleic acid sequence in a sample suspected of containing the same. The assay utilizes nucleic acid matrices to facilitate reactions which detect the presence of a nucleic acid sequence through formation of a hybridized or bound nucleic acid structure having a portion capable of disassociating therefrom through the action of a structure specific cleaving enzyme.
Claims27
A nucleic acid assay for detecting the presence of a specific sample nucleic acid sequence in a sample suspected of the containing the same, said nucleic acid assay comprising:
- (a) a matrix comprising at least one first site for receiving an invader oligonucleotide and at least one second site for receiving a probe oligonucleotide;
- (b) at least one invader oligonucleotide for attaching to the first site of the matrix, said invader oligonucleotide having an invader nucleic acid sequence for binding to a first portion of the sample nucleic acid sequence;
- (c) at least one probe oligonucleotide for attaching to the second site of the matrix, said probe oligonucleotide having a first probe nucleotide portion for binding to a second portion of the sample nucleic acid sequence and a second probe nucleotide portion which does not bind to the sample nucleic acid sequence;
- (d) a first disassociating agent for disassociating the second probe nucleotide portion of the probe oligonucleotide from the first probe nucleotide portion upon the concurrent binding of the invader nucleic acid sequence of an invader oligonucleotide to the first portion of the sample nucleic acid sequence and the first probe nucleotide portion for binding to the second portion of the sample nucleic acid sequence;
- and
- (e) detection means for detecting the degree to which the second probe nucleotide portion of the probe oligonucleotide has disassociated from the first probe nucleotide portion thereof.
A method of detecting the presence of a specific sample nucleic acid sequence in a sample suspected of containing the same, said method comprising the steps of:
- contacting the sample with a matrix comprising at least one first site with an invader oligonucleotide having an invader nucleic acid sequence for binding to a first portion of the specific sample nucleic acid sequence, and at least one second site with a probe oligonucleotide having a first probe nucleotide portion for binding to a second portion of the specific sample nucleic acid sequence and a second probe nucleotide portion which does not bind to the specific sample nucleic acid sequence to yield a sample-matrix mixture;
- treating the sample-matrix mixture at a temperature and for a time sufficient to induce the invader oligonucleotide to bind to the first portion of the specific sample nucleic acid sequence and to induce the first probe nucleotide portion of the probe oligonucleotide to bind to the second portion of the specific sample nucleic acid sequence to yield a hybridization complex;
- contacting the hybridization complex with a disassociating agent wherein the disassociating agent disassociates the second probe nucleotide portion from the first probe nucleotide portion thereof;
- and
- detecting the degree to which the second probe nucleotide portion of the probe oligonucleotide has disassociated from the first probe nucleotide portion thereof.
Dependencies are inferred from the claim text.