Patent ID: 6593091
Filing Date: 2003-07-15
Classification: C12Q

Abstract:
A method for detecting a polynucleotide target in a sample, the method comprising the steps of:a) forming a mixture by combining: (i) a sample known to contain or suspected of containing the polynucleotide target; (ii) a first probe complementary to the polynucleotide target, the first probe comprising a first fluorescent donor or fluorescent acceptor, a first segment and a second segment; (iii) a second probe partially complementary to the first probe, the second probe comprising a second fluorescent donor or fluorescent acceptor, a first segment and a second segment, wherein the second probe competes with the polynucleotide target for binding to the first probe and the first probe preferentially binds to the polynucleotide target rather than to the second probe, wherein the first segments of the first and second probes are complementary and the second segments of the first and second probes are not complementary, and wherein both the first and second segments of at least the first probe are complementary to the polynucleotide target, wherein the first fluorescent donor or acceptor and the second fluorescent donor or acceptor are a fluorescent donor/acceptor pair capable of fluorescence resonance energy transfer with each other in response to activation of the fluorescent donor by light of a predetermined wavelength or band of wavelengths, and wherein upon hybridization of the first probe to the second probe the fluorescent donor and the fluorescent acceptor are spaced apart from each other a distance such that they are capable of fluorescence resonance energy transfer in response to activation of the fluorescent donor by light of the predetermined wavelength; b) activating the fluorescent donor in the mixture with light of the predetermined wavelength or band of wavelengths; and c) detecting light emitted by the fluorescent donor, the fluorescent acceptor, or both fluorescent donor and acceptor.