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Associated Analysis of DNA Methylation for Cancer Detection Using CCP-Based FRET Technique
journal contribution
posted on 2014-01-07, 00:00 authored by Jiangyan Zhang, Baoling Xing, Jinzhao Song, Feng Zhang, Chenyao Nie, Lian Jiao, Libing Liu, Fengting Lv, Shu WangThis
paper describes an associated analysis method of DNA methylation
for the detection of cancer using an optically amplifying cationic
conjugated polymer (CCP, poly{(1,4-phenylene)-2,7-[9,9-bis(6′-N,N,N-trimethyl ammonium)-hexyl
fluorene] dibromide)}. Genomic DNA is digested by methylation-sensitive
restriction endonuclease, followed by PCR amplification to incorporate
fluorescein-labeled dNTP. Only methylated DNA can be amplified by
PCR, and the methylation level is detected through fluorescence resonance
energy transfer (FRET) between CCP and fluorescein that is incorporated
into the PCR product. The methylation levels of RASSF1A, OPCML, and HOXA9 promoters of
35 ovarian cancer samples and 11 normal samples were assayed. In accordance
with the degree of methylation levels, they are clustered to three
sections and assigned a value. Through an associated analysis, we
acquired a threshold for cancer detection with a sensitivity of 85.7%.
The assay takes about 20 h to obtain the detection results and shows
great potential as a useful tool for diagnostic and screening of cancer.