Patent ID: 7838225

Claim:
A method for detecting a target nucleic acid sequence in a sample comprising, mixing an upstream primer, a downstream probe, wherein the downstream probe comprises a 5′ region and a 3′ region, wherein the 3′ region is at least partially complementary to the target nucleic acid and the 5′ region forms a flap, a reverse transcriptase, a cleavage agent and a target nucleic acid under conditions which are permissive for the steps of (i) annealing of the upstream primer and downstream probe, (ii) extending the upstream primer wherein the reverse transcriptase synthesizes a primer extension product and forms a cleavage structure with the downstream probe, and (iii) cleaving the cleavage structure with the cleavage agent to generate a detectable signal (iv) detecting and/or measuring said detectable signal, wherein detecting and/or measuring of said detectable signal is indicative of the presence of said target nucleic acid.