Fluorescence resonance energy transfer dye-labeled probe for fluorescence-enhanced DNA detection: An effective strategy to greatly improve discrimination ability toward single-base mismatch

In this article, we report on the first use of fluorescence resonance energy transfer (FRET) dye-labeled probe for fluorescence resonance enhanced DNA detection to greatly improve discrimination ability toward single-base mismatch using conjugation polymer poly(p-phenylenediamine) nanobelts (PNs) as a sensing platform. The suggested FRET dye-labeled probe contains a 5-carboxyfluorescein (FAM) group at 5′ end of the oligomer as a donor and a 6-carboxy-X-rhodamine (ROX) attached to a modified cytosine (C) base as an acceptor, which were separated by three bases. The general concept used in this DNA assay is based on adsorption of the FRET dye-labeled single-stranded DNA (ssDNA) probe by PN, which is accompanied by substantial fluorescence quenching and disappearance of FRET. The subsequent specific hybridization with its target forms a double-stranded DNA (dsDNA), resulting in desorption of the hybridized duplex from PN surface accompanied by reoccurrence of FRET and fluorescence recovery. It suggests that the discrimination ability of this FRET probe based system toward single-base mismatch is about 5.2 times that of the system based on single dye-labeled probe based system.

► FRET dye-labeled probe improves discrimination ability to single-base mismatch.
► It solves the low discrimination ability issue with the nano/microstructure platforms.
► This fluorescent sensing system holds great potential for practical mismatch detection.