Methods for detecting a target nucleotide sequence in a sample utilising a nuclease-aptamer complex

摘要

The present invention relates to methods for detecting a target nucleotide sequence in a sample. More particularly, the present invention relates to methods for detecting a target nucleotide sequence in a sample which utilise a nuclease-aptamer complex. The present invention also provides nuclease-binding aptamers, nuclease-aptamer complexes and linker molecules that may be used in accordance with the methods of the present invention.

主权项

1. A method for detecting a target nucleotide sequence in a sample, the method comprising:
providing a nuclease-aptamer complex as set forth in FIG. 1, the complex comprising a nuclease bound to an aptamer, wherein binding of the aptamer to the nuclease inhibits the activity of the nuclease; providing a linker nucleic acid comprising a first portion which can hybridise with the target nucleotide sequence if it is present in the sample, and a second portion which can hybridise with the aptamer when the first portion of the linker nucleic acid is hybridised to the target nucleotide sequence, wherein hybridisation between the aptamer and the second portion of the linker nucleic acid modifies, reduces or eliminates binding between the aptamer and the nuclease, and wherein the linker nucleic acid comprises a stem loop structure and at least a portion of the first portion of the linker is comprised within a loop of the stem-loop structure and at least a portion of the second portion of the linker is comprised within a stem of the stem-loop structure; applying the nuclease-aptamer complex to the sample; applying the linker nucleic acid to the sample; allowing the first portion of the linker nucleic acid to hybridise with the target nucleotide sequence if it is present in the sample; allowing the second portion of the linker nucleic acid to hybridise with the aptamer when the first portion of the linker nucleic acid hybridises with the target nucleotide sequence; and detecting the activity of the nuclease, wherein increased nuclease activity in the sample is indicative of the presence of the target nucleotide sequence in the sample.