TY - JOUR
T1 - A novel nucleic acid fluorescent sensing platform based on nanostructured films of intrinsically conducting polymers
AU - Pedro, Graciela C.
AU - Gorza, Filipe D.S.
AU - da Silva, Romário J.
AU - do Nascimento, Kamila T.O.
AU - Medina-Llamas, Juan C.
AU - Chávez-Guajardo, Alicia E.
AU - Alcaraz-Espinoza, José J.
AU - de Melo, Celso P.
N1 - Publisher Copyright:
© 2018
PY - 2019/1/24
Y1 - 2019/1/24
N2 - 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 ds-DNA 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.
AB - 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 ds-DNA 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.
KW - Conducting polymers
KW - Fluorescent detection
KW - Molecular diagnosis
KW - Nanostructured films
KW - Test strips
UR - http://www.scopus.com/inward/record.url?scp=85055030132&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2018.10.010
DO - 10.1016/j.aca.2018.10.010
M3 - Artículo
C2 - 30567653
SN - 0003-2670
VL - 1047
SP - 214
EP - 224
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
ER -