title: Associated analysis of DNA methylation for cancer detection using CCP-based FRET technique.
abstract: This 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.
authors: Beijing National Laboratory for Molecular Sciences, Key Laboratory of Organic Solids, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, P. R. China.
authors: Jiangyan
authors: J
authors: Zhang
authors: Baoling
authors: B
authors: Xing
authors: Jinzhao
authors: J
authors: Song
authors: Feng
authors: F
authors: Zhang
authors: Chenyao
authors: C
authors: Nie
authors: Lian
authors: L
authors: Jiao
authors: Libing
authors: L
authors: Liu
authors: Fengting
authors: F
authors: Lv
authors: Shu
authors: S
authors: Wang
mesh_terms: D002412:Cations; D019175:DNA Methylation / Q000235:genetics*; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D009369:Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000378:metabolism
chemical_list: D002412:Cations