Rapid Electrochemical Assessment of Tumor Suppressor Gene Methylations in Raw Human Serum and Tumor Cells and Tissues Using Immunomagnetic Beads and Selective DNA Hybridization.
Angew Chem Int Ed Engl
; 57(27): 8194-8198, 2018 07 02.
Article
in En
| MEDLINE
| ID: mdl-29744991
ABSTRACT
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 500â
pm and a limit of detection of 1.2â
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.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
DNA Modification Methylases
/
DNA Methylation
/
Tumor Suppressor Proteins
/
5-Methylcytosine
/
DNA Repair Enzymes
/
Electrochemical Techniques
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Year:
2018
Type:
Article