期刊
ACS SENSORS
卷 4, 期 1, 页码 227-234出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.8b01339
关键词
global and gene-specific DNA methylation; 5-mC; 5-hmC; electrochemical platforms; cancer; human serum; paraffin-embedded colorectal tissues
资金
- Spanish Ministerio de Economia y Competitividad [CTQ2015-70023-R, CTQ2015-64402-C2-1-R]
- AES-ISCIII [PI17CIII/00045 (UFIECPY 124/18)]
- NANO-AVANSENS Program from the Comunidad de Madrid [S2013/MT-3029]
- Spanish Ministerio de Economia y Competitividad
- Universidad Complutense de Madrid
This paper reports the preparation of versatile electrochemical biosensing platforms for the simple, rapid, and PCR-independent detection of the most frequent DNA methylation marks (5-methylcytosine, 5-mC, and/or 5-hydroxymethylcytosine, 5-hmC) both at global and gene-specific levels. The implemented strategies, relying on the smart coupling of immuno-magnetic beads (MBs), specific DNA probes and amperometric detection at screen-printed carbon electrodes (SPCEs), provided sensitive and selective determination of the target methylated DNAs in less than 90 min with a great reproducibility and demonstrated feasibility for the simultaneous detection of the same or different cytosine epimarks both at global level and in different loci of the same gene or in different genes. The bioplatforms were applied to determine global methylation events in paraffin-embedded colorectal tissues genes in genomic DNA extracted from cancer cells and paraffin-embedded colorectal tissues, and in serum without previous DNA extraction from cancer patients.
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