Georgetown University

Mark Danielsen

Eugene A Davidson

Bolor Tumurpurev

Kenneth L Dretchen

Berenice Alfonso

Joel Credle

Traci K Pals

The present invention relates to a novel method for detecting a target polynucleotide having a target sequence, comprising hybridizing the target polynucleotide with a probe to form a hybrid; exposing the hybrid to a 5' exonuclease so that the probe in the hybrid is digested and the target polynucleotide is dissociated from the digested probe; repeating the hybridization step and the digestion step; and detecting the digested probes. The presence of the digested probes indicates the presence of the target polynucleotide.

Exonuclease cycling assay

The present invention relates to a novel method for detecting a target polynucleotide having a target sequence, comprising (a) exposing the target polynucleotide to an initiating oligonucleotide; (b) extending the initiating oligonucleotide with an extended sequence complementary to the target sequence; (c) ligating the initiating oligonucleotide sequence with the extended sequence to form a circular oligonucleotide having a nicking endonuclease (NE) recognition/cutting sequence; (d) exposing the circular oligonucleotide to a DNA polymerase and a DNA synthesis primer to synthesize DNA having a NE recognition sequence; (e) exposing the synthesized DNA to a probe having the NE recognition/cutting sequence to form a double stranded DNA having a full NE site; (f) exposing the double stranded DNA to a nicking endonuclease (NE) to cleave the probe; and (g) detecting the cleaved probe. The presence of the cleaved probe indicates the presence of the target polynucleotide.

Polymerase driven NESA

A method for detecting the presence of a target nucleotide sequence in a sample of DNA is described herein in which a test sample comprising single stranded DNA is exposed to a DNA probe and a nicking endonuclease under conditions that would permit sequence-specific hybridization of the probe to a complementary target sequence. The probe comprises a sequence complementary to the target sequence to be detected and this sequence also includes a recognition sequence for the nicking endonuclease. If the sample contains the target sequence, the probe hybridizes to the target and is cleaved by the nicking endonuclease, which leaves the target intact. Observing the presence of probe cleaved by the nicking endonuclease indicates the presence of the target nucleotide sequence in the sample of DNA.

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Germantown, MD, United States of America

Mark Danielsen