Method for detecting nucleic acid target sequences involving in vitro transcription from an rna polymerase promoter

Abstract

Disclosed is a complex formed by a hybridization reaction comprising four nucleic acid molecules; the complex comprising a target nucleic acid molecule and first, second and third nucleic acid probe molecules; wherein the first probe comprises a foot region which is complementary to a first portion of the target and is hybridized thereto, and an arm region which is substantially non-complementary to the target; the second probe comprises a foot region which is complementary to a second portion of the target, such that the foot region of the second probe is hybridized to the target adjacent or substantially adjacent to the foot region of the first probe, the second probe also comprising an arm region which is substantially non-complementary to the target but which is complementary and hybridized to the arm region of the first probe; the third probe being complementary, at least in part, to a portion of the arm region of the first probe, such that third probe is hybridized to the arm region of the first probe adjacent or substantially adjacent to the second probe; and wherein formation of the complex creates a functional double-stranded RNA polymerase promoter, one strand of the promoter being provided by the first probe, and the other strand being provided jointly by the second probe and by the third probe; and a method of detecting a target nucleic acid sequence of interest which method involves the formation of the complex.