2026-07-21T07:17:47Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/91492025-05-05T07:26:18Zcom_20.500.12105_15322com_20.500.12105_2051com_20.500.12105_2052col_20.500.12105_16963col_20.500.12105_19617
Repisalud
author
Povedano, Eloy
author
Vargas, Eva
author
Ruiz-Valdepeñas Montiel, Víctor
author
Torrente-Rodríguez, Rebeca M
author
Pedrero, María
author
Barderas Manchado, Rodrigo
author
Segundo-Acosta, Pablo San
author
Peláez-García, Alberto
author
Mendiola, Marta
author
Hardisson, David
author
Campuzano, Susana
author
Pingarrón, José M
funder
Ministerio de Economía y Competitividad (España)
funder
Comunidad de Madrid (España)
funder
Complutense University of Madrid (España)
2020-02-26T15:27:24Z
2020-02-26T15:27:24Z
2018
Sci Rep. 2018 Apr 23;8(1):6418.
10.1038/s41598-018-24902-1
2045-2322
2045-2322
Scientific reports
29686400
http://hdl.handle.net/20.500.12105/9149
This paper describes two different electrochemical affinity biosensing approaches for the simple, fast and bisulfite and PCR-free quantification of 5-methylated cytosines (5-mC) in DNA using the anti-5-mC antibody as biorecognition element. One of the biosensing approaches used the anti-5-mC as capture bioreceptor and a sandwich type immunoassay, while the other one involved the use of a specific DNA probe and the anti-5-mC as a detector bioreceptor of the captured methylated DNA. Both strategies, named for simplicity in the text as immunosensor and DNA sensor, respectively, were implemented on the surface of magnetic microparticles and the transduction was accomplished by amperometry at screen-printed carbon electrodes by means of the hydrogen peroxide/hydroquinone system. The resulting amperometric biosensors demonstrated reproducibility throughout the entire protocol, sensitive determination with no need for using amplification strategies, and competitiveness with the conventional enzyme-linked immunosorbent assay methodology and the few electrochemical biosensors reported so far in terms of simplicity, sensitivity and assay time. The DNA sensor exhibited higher sensitivity and allowed the detection of the gene-specific methylations conversely to the immunosensor, which detected global DNA methylation. In addition, the DNA sensor demonstrated successful applicability for 1 h-analysis of specific methylation in two relevant tumor suppressor genes in spiked biological fluids and in genomic DNA extracted from human glioblastoma cells.
eng
Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments
research article
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ElectrochemicalAffinityBiosensorsFor_2018.pdf
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ElectrochemicalAffinityBiosensorsFor_2018.pdf.txt