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

00925njm 22002777a 4500 dc Torrente Rodríguez, Rebeca Magnolia author Campuzano Ruiz, Susana author Ruiz Valdepeñas Montiel, Víctor author Montoya Miñano, Juan José author Pingarrón Carrazón, José Manuel author 2016 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. 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. 0956-5663 10.1016/j.bios.2016.07.003 https://hdl.handle.net/20.500.14352/96114 https://doi.org/10.1016/j.bios.2016.07.003 Sensitive electrochemical determination of miRNAs based on a sandwich assay onto magnetic microcarriers and hybridization chain reaction amplification