Patent ID: 6696255
Filing Date: 2004-02-24
Classification: C12Q,Y10T

Abstract:
A method for detecting a target nucleotide sequence in a sample comprising the steps of:a) providing an oligonucleotide probe comprising a nucleotide sequence that forms a hairpin structure having a stem region comprising a double stranded segment under suitable conditions, wherein the double stranded segment is formed between two complementary nucleotide sequences, and wherein at least a portion of at least one of the nucleotide sequences located within the double stranded segment is complementary to the target nucleotide sequence to be detected; b) contacting the probe provided in step a) with a sample containing or suspected of containing the target nucleotide sequence under conditions that favor intermolecular hybridization between the probe and the target nucleotide sequence over intramolecular hybridization of the probe itself, whereby the contacting breaks the intramolecular hybridization of the probe itself by dissociating the double stranded segment of the probe and forms a first intermolecular hybrid between the target and a region consisting of at least a portion of one strand of said stem that is complementary to the target nucleotide sequence; and c) assessing the first intermolecular hybrid formed in step b), wherein the assessing is effected by addition of a secondary probe that specifically hybridizes with at least a portion of the first intermolecular hybrid, thereby forming a secondary intermolecular hybrid formed between the secondary probe and the first intermolecular hybrid wherein the detection of the secondary intermolecular hybrid indicates the presence and/or amount of the target nucleotide sequence in the sample.