Patent ID: 7785784
Filing Date: 2010-08-31
Classification: C12Q

Abstract:
1. A method of detecting a target oligonucleotide in a sample, comprising the steps of: (a) contacting the sample with a first polynucleotide that is complementary to a first portion of the target oligonucleotide, so as to allow hybridization of the first polynucleotide with the target oligonucleotide, said first polynucleotide is joined to a tag, wherein the target oligonucleotide is an anti-VEGF aptamer having the structure: (b) contacting the sample with a second polynucleotide that is complementary to an adjacent and non-overlapping portion of the target oligonucleotide so as to allow hybridization of the second polynucleotide with the target oligonucleotide thereby forming a hybrid complex, said second polynucleotide comprises an amplifiable sequence that is substantially non-complementary to the target oligonucleotide; and (c) detecting the amplifiable sequence of the second polynucleotide in the hybrid complex by an amplification step, wherein the amplifiable sequence is detected using a fluorescent probe selected from the group consisting of a fluorescent probe comprising a nucleic acid having the sequence AAG GAA CTC TTC TGG AAT GA (SEQ ID NO: 8) and a fluorescent probe having the structure: 6FAM-AAG GAA CTC TTC TGG AAT GA-MGBNFQ (SEQ ID NO: 9), whereby, detection of the amplifiable sequence indicates that the target oligonucleotide is present in the sample.