| 摘要 |
<p>The present invention relates to methods and compositions for detecting a target nucleotide sequence in a sample. In particular, the methods and compositions of the present invention employ a reporter enzyme tethered to a solid support via a tether nucleotide sequence. A disclosed method for detecting a target nucleotide sequence comprises providing a linker nucleotide sequence comprising a probe sequence that can hybridize to the target sequence and a trigger which can hybridize to the tether nucleotide sequence when the probe sequence binds to the target. The linker nucleotide can have a stem-loop structure. When the trigger hybridizes to the tether nucleotide sequence, the tether is able to be digested by a nuclease, releasing the reporter enzyme from the solid support. The activity of the released reporter enzyme on a spatially or temporally separated substrate generates a detectable signal. In some embodiments, the reporter enzyme releases a feedback trigger that can hybridize to the tether nucleotide sequence and release further reporter enzymes, amplifying the signal via a feedback loop.</p> |
