Patent ID: 6379888
Filing Date: 2002-04-30
Classification: C12Q,Y10T

Abstract:
A method for detecting the presence of a nucleic acid target sequence comprising:a) hybridizing to the target sequence a signal primer comprising i) a reporter probe which in the absence of hybridization to a complementary sequence assumes a conformational structure which brings a fluorescent donor/quencher dye pair linked thereto into sufficiently close spatial proximity to quench donor fluorescence, and, ii) an adapter oligonucleotide hybridized to the reporter probe such that the signal primer comprises an intermolecularly base-paired portion and a single-stranded target binding sequence, wherein quenching of donor fluorescence in the absence of hybridization of the adapter oligonucleotide and the reporter probe is greater than quenching of donor fluorescence when the adapter oligonucleotide and the reporter probe are hybridized;b) in a primer extension reaction, synthesizing a strand complementary to the adapter oligonucleotide, whereby the reporter probe is separated from the adapter oligonucleotide and quenching of donor fluorescence is increased, and; c) detecting a change in a fluorescence parameter associated with increased quenching as an indication of the presence of the target sequence.