Homogeneous diagnostic assay method utilizing simultaneous target and signal amplification

摘要

The present invention relates to a method, kit and reagent composition for determining the presence of a target nucleic acid in a sample using a two-stage target cycling reaction. During stage 1 of the reaction, the target nucleic acid hybridizes with a probe nucleic acid to effectuate release of an activator into the assay medium. During stage 2 of the reaction, a target analog-anchor complex is cleaved by the activator, which accomplishes the dual purpose of releasing the target analog into the assay medium, and the initiation of signal generation. The released target analog then restarts the cyclic reaction by binding to a second probe, which effectuates release of a second activator, and so on. This cyclic reaction gives rise to an increase in the amount of signal generated from a single target nucleic acid molecule in the sample, which greatly enhances the level of target detection that can be expected.