Patent ID: 8247171

Claim:
A method for detection of the presence of a target polynucleotide in a sample, said method comprising a) providing a mixture of said sample, nucleic acid amplification reagents and a target binding agent, said target binding agent being capable of assuming a first position where said target binding agent is not bound to said target polynucleotide and a second position where said target binding agent is bound to said target polynucleotide, and a signal is measurable from said mixture, the intensity of said signal depending on the proportions of target binding agent being in said first and second positions; b) exposing said mixture to conditions that allow nucleic acid amplification to occur in said mixture and allowing said amplification to complete; c) exposing said mixture to a first condition resulting in that said target binding agent is in said first position; d) exposing said mixture to a second condition where said target binding agent is capable of assuming the second position; e) measuring at least once, after completion of said nucleic acid amplification and after said mixture has been exposed to said first condition, said signal of the mixture at a first time when less than in the step f) of said target binding agent present in said mixture has assumed the second position; f) measuring at least once, after completion of said nucleic acid amplification and after said mixture has been exposed to said second condition, said signal of the mixture at a second time when, if the sample contains said target polynucleotide, more than in step e) of said target binding agent has assumed the second position; and g) determining the ratio of the measurement results obtained in steps e) and f) and comparing it to a pre-determined cut-off value that is characteristic for said mixture of sample and target binding agent to determine whether said target polynucleotide is present in the sample.