4.8 Article

Rapid Electrochemical Assessment of Tumor Suppressor Gene Methylations in Raw Human Serum and Tumor Cells and Tissues Using Immunomagnetic Beads and Selective DNA Hybridization

Journal

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 57, Issue 27, Pages 8194-8198

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201804339

Keywords

biofluids; brain tissues; dual detection; immuno-DNA sensors; methylated DNA

Funding

  1. Spanish Ministerio de Economia y Competitividad [CTQ2015-70023-R, CTQ2015-64402-C2-1-R, SAF2014-53209-R]
  2. AESI [PI17CIII/00045]
  3. NANOAVANSENS Program from the Comunidad de Madrid [S2013/MT-3029]
  4. Spanish Ministerio de Economia y Competitividad
  5. Spanish Ministerio de Educacion, Cultura y Deporte
  6. Universidad Complutense de Madrid

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We report a rapid and sensitive electrochemical strategy for the detection of gene-specific 5-methylcytosine DNA methylation. Magnetic beads (MBs) modified with an antibody for 5-methylcytosines (5-mC) are used for the capture of any 5-mC methylated single-stranded (ss)DNA sequence. A flanking region next to the 5-mCs of the captured methylated ssDNA is recognized by hybridization with a synthetic biotinylated DNA sequence. Amperometric transduction at disposable screen-printed carbon electrodes (SPCEs) is employed. The developed biosensor has a dynamic range from 3.9 to 500 pm and a limit of detection of 1.2 pm for the methylated synthetic sequence of the tumor suppressor gene O-6-methylguanine-DNA methyltransferase (MGMT) promoter region. The method is applied in the 45-min analysis of specific methylation in the MGMT promoter region directly in raw spiked human serum samples and in genomic DNA extracted from U-87 glioblastoma cells and paraffin-embedded brain tumor tissues without any amplification and pretreatment step.

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