Rapid detection of a dengue virus RNA sequence with single molecule sensitivity using tandem toehold-mediated displacement reactions

Abstract

A DNA tetrahedron having a single-strand DNA edge was used to detect a dengue virus RNA sequence. Novel tandem toehold-mediated displacement reactions (tTMDR) were developed to amplify the fluorescence signal from the DNA tetrahedron. Using an excess of the DNA tetrahedron each target RNA was recycled about 10³ times during the tTMDR process. This amplification process was used for the sensitive detection of dengue virus RNA in this study. As few as 6 copies of RNA per sample could be detected using a photon count technique with single molecule sensitivity.