Capture based nucleic acid detection

摘要

The present invention relates to methods and kits useful for the detection of a target nucleotide sequence in a sample. In general, the methods of the present invention are predicated on the target-mediated capture of a nuclease into a complex wherein the extent of complex formation, as measured by nuclease activity, is positively correlated with the presence of the target nucleic acid in the sample.

主权项

1. A method for detecting a target nucleic acid in a sample, the method comprising forming a reaction mix comprising:
the sample; a nuclease; a solid substrate; a probe nucleic acid which is hybridisable under the conditions of the method to a first portion of the target nucleic acid, wherein the probe nucleic acid is immobilisable on the solid substrate; and a capture structure comprising a capture portion which is able to capture the nuclease and a targeting portion comprising a nucleotide sequence which is hybridisable under the conditions of the method to a second portion of the target nucleic acid; wherein the presence of the target nucleic acid in the sample allows the formation of a complex on the solid substrate wherein the complex comprises the probe nucleic acid, the target nucleic acid, the capture structure and a captured nuclease; and determining the extent of complex formation comprising measuring a residual nuclease activity in the reaction mix, wherein the extent of the residual nuclease activity is negatively correlated with the extent of complex formation, and wherein the extent of complex formation is positively correlated with the presence of the target nucleic acid in the sample.