Versatile Electroanalytical Bioplatforms for Simultaneous Determination of Cancer-Related DNA 5-Methyl- and 5-Hydroxymethyl-Cytosines at Global and Gene-Specific Levels in Human Serum and Tissues.
Eloy PovedanoVíctor Ruiz-Valdepeñas MontielAlejandro ValverdeFernando Navarro-VillosladaPaloma Yáñez-SedeñoMaría PedreroAna Montero-CalleRodrigo BarderasAlberto Peláez-GarcíaMarta MendiolaDavid HardissonJaime FeliúJordi CampsElisabet Rodríguez-TomàsJorge JovenMeritxell ArenasSusana CampuzanoJosé Manuel PingarrónPublished in: ACS sensors (2018)
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 and specific methylation at promoters of tumor suppressor genes in genomic DNA extracted from cancer cells and paraffin-embedded colorectal tissues, and in serum without previous DNA extraction from cancer patients.
Keyphrases
- genome wide
- dna methylation
- circulating tumor
- copy number
- single molecule
- gene expression
- loop mediated isothermal amplification
- label free
- genome wide identification
- simultaneous determination
- cell free
- molecularly imprinted
- real time pcr
- nucleic acid
- solid phase extraction
- liquid chromatography tandem mass spectrometry
- high performance liquid chromatography
- gold nanoparticles
- small molecule
- genome wide analysis
- high throughput
- emergency department
- mass spectrometry
- transcription factor
- high resolution
- single cell
- ms ms
- sensitive detection
- carbon nanotubes
- adverse drug
- living cells
- transition metal