Patent ID: 8916349

Claim:
A method of detecting the presence or absence of a target nucleic acid in a sample, comprising: a) preparing a sample suspected of containing a target nucleic acid; b) preparing a reaction mixture comprising the sample, first and second oligonucleotide probes, and a sequence specific endonuclease, wherein the first and second oligonucleotide probes each comprise a first sequence and a second sequence, wherein the first sequences of the first and second oligonucleotide probes are complementary with non-overlapping sequences of the target nucleic acid, and wherein the second sequences of each of the first and second oligonucleotide probes are complementary with each other, thus forming a duplex restriction site recognized by the sequence specific endonuclease, and wherein at least one of the first and second oligonucleotide probes comprises a detectable label positioned at one side of the restriction site, and a quenching moiety positioned at the other side of the restriction site, thus allowing the first sequences of the first and second oligonucleotide probes to selectively hybridize with the non-overlapping sequences of the nucleic acid of the target nucleic acid and the second sequences of the first and second oligonucleotide probes to selectively hybridize with each other and thereby form the duplex restriction site, such that, if the target nucleic acid is present in the sample, the sequence specific endonuclease cleaves the duplex at the restriction site, allowing for detection of the label, and denaturing the remaining portion of the oligonucleotide probes from the nucleic acid of the target organism and from each other.