2026-06-28T09:49:46Zhttps://docta.ucm.es/rest/oai/requestoai:docta.ucm.es:20.500.14352/126482023-08-27T07:18:00Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense
author
Vargas Orgaz, Eva
author
Povedano Muñumel, Eloy
author
Ruiz Valdepeñas Montiel, Víctor
author
Torrente Rodríguez, Rebeca Magnolia
author
Zouari, Mohamed
author
Montoya Miñano, Juan José
author
Raouafi, Noureddine
author
Campuzano Ruiz, Susana
author
Pingarrón Carrazón, José Manuel
2023-06-17T12:37:32Z
2023-06-17T12:37:32Z
2018-03-15
1424-8220
10.3390/s18030863
https://hdl.handle.net/20.500.14352/12648
https://doi.org/10.3390/s18030863
https://www.mdpi.com/1424-8220/18/3/863
This work reports an amperometric biosensor for the determination of miRNA-21, a relevant oncogene. The methodology involves a competitive DNA-target miRNA hybridization assay performed on the surface of magnetic microbeads (MBs) and amperometric transduction at screen-printed carbon electrodes (SPCEs). The target miRNA competes with a synthetic fluorescein isothiocyanate (FITC)-modified miRNA with an identical sequence for hybridization with a biotinylated and complementary DNA probe (b-Cp) immobilized on the surface of streptavidin-modified MBs (b-Cp-MBs). Upon labeling, the FITC-modified miRNA attached to the MBs with horseradish peroxidase (HRP)-conjugated anti-FITC Fab fragments and magnetic capturing of the MBs onto the working electrode surface of SPCEs. The cathodic current measured at −0.20 V (versus the Ag pseudo-reference electrode) was demonstrated to be inversely proportional to the concentration of the target miRNA. This convenient biosensing method provided a linear range between 0.7 and 10.0 nM and a limit of detection (LOD) of 0.2 nM (5 fmol in 25 μL of sample) for the synthetic target miRNA without any amplification step. An acceptable selectivity towards single-base mismatched oligonucleotides, a high storage stability of the b-Cp-MBs, and usefulness for the accurate determination of miRNA-21 in raw total RNA (RNAt) extracted from breast cancer cells (MCF-7) were demonstrated.
eng
Atribución 3.0 España
Single-Step Incubation Determination of miRNAs in Cancer Cells Using an Amperometric Biosensor Based on Competitive Hybridization onto Magnetic Beads
journal article
URL
https://docta.ucm.es/bitstreams/7c7025a0-ddee-44be-8e7b-ab1d2b17757f/download
File
MD5
b3376e773a3a9d37b50202e97bc2cb1c
1811755
application/pdf
sensors-18-00863.pdf
URL
https://docta.ucm.es/bitstreams/de07ca1f-d3ad-42b7-8030-c072184b7c98/download
File
MD5
cd8d3a950a4cb751245cdddcfcbabfff
62747
text/plain
sensors-18-00863.pdf.txt