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oai:docta.ucm.es:20.500.14352/961142025-01-09T19:37:55Zcom_20.500.14352_14col_20.500.14352_15

Sensitive electrochemical determination of miRNAs based on a sandwich assay onto magnetic microcarriers and hybridization chain reaction amplification Torrente Rodríguez, Rebeca Magnolia Campuzano Ruiz, Susana Ruiz Valdepeñas Montiel, Víctor Montoya Miñano, Juan José Pingarrón Carrazón, José Manuel 543 MiRNA-21 Magnetic beads Hybridization chain reaction SPCEs Tumor cell Human tissues Química analítica (Química) 2301 Química Analítica A novel electrochemical approach for determination of miRNAs involving a sandwich hybridization assay onto streptavidin-magnetic beads (Strep-MBs), hybridization chain reaction (HCR) amplification and amperometric detection at disposable screen-printed carbon electrodes is reported. Using miRNA-21 as the target analyte, a dynamic linear range from 0.2 to 5.0 nM with a 60 pM (1.5 fmol in 25 μL) detection limit was obtained. The achieved sensitivity is 24-fold higher than a non-HCR amplification approach involving conventional sandwich type assay onto MBs. Moreover, the whole assay time lasted 1 h 45 min which is remarkably shorter than other reported methodologies. The methodology exhibited full selectivity against other non-complementary miRNAs as well as an acceptable discrimination between homologous miRNA family members. The applicability of this novel approach was demonstrated by determining mature miRNA-21 in total RNA (RNAt) extracted from tumor cells and human tissues. Ministerio de Economía y Competitividad (España) Comunidad de Madrid CANAAN Research & Investment S.L. (España) Universidad Complutense de Madrid Depto. de Química Analítica Fac. de Ciencias Químicas TRUE pub 2024-01-29T13:42:15Z 2024-01-29T13:42:15Z 2016 journal article VoR https://hdl.handle.net/20.500.14352/96114 0956-5663 10.1016/j.bios.2016.07.003 eng CTQ2015-64402-C2-1-R S2013/ MT-3029 Art.83 Project Torrente-Rodríguez, R.M., et al., Biosensors and Bioelectronics (2016), http://dx.doi.org/10.1016/j. bios.2016.07.003i Torrente-Rodríguez, R. M., et al. «Sensitive Electrochemical Determination of miRNAs Based on a Sandwich Assay onto Magnetic Microcarriers and Hybridization Chain Reaction Amplification». Biosensors and Bioelectronics, vol. 86, diciembre de 2016, pp. 516-21. https://doi.org/10.1016/j.bios.2016.07.003. restricted access application/pdf Elsevier