A novel nucleic acid fluorescent sensing platform based on nanostructured films of intrinsically conducting polymers

摘要

When fluorophores attach to nanostructured films of intrinsically conducting polymers (ICPs), a quenching of their fluorescence may occur. We have exploited these characteristics for the development of polymeric films that can be used in a simple and efficient molecular diagnosis protocol based on the selective detection of nucleic acids. Our procedure rests on the fact that the fluorescence of 6-carboxyfluorescein-labeled single-stranded DNA (FAM-ssDNA) probes is quenched upon their immobilization on nanostructured ICP - polypyrrole (PPY) and polyaniline (PANI) - films deposited on polyethylene terephthalate (PET) substrates. Hybridization occurs whenever a sample with the complementary sequence is brought in contact with the immobilized probe, with the newly formed dsDNA chains detaching from the flexible polymeric film and causing the restoration of the fluorescence. This sensing system exhibits a low background signal that depends on both the thickness and hydrophobicity of the films. As a model system, we used a FAM-ssDNA probe specific for the Leishmania infantum parasite. The results confirm this procedure as a simple, fast and highly sensitive scheme for the recognition of the target DNA, with a detection limit of the 1.1 nM and 1.3 nM for the PPY/PET and PANI/PET films, respectively. In addition, this biosensor has excellent stability and exhibits a good and reproducible performance even when used for the direct detection of ssDNA in relatively complex biological samples. (c) 2018 Published by Elsevier B.V.