title: Mass Spectrometry Based Ultrasensitive DNA Methylation Profiling Using Target Fragmentation Assay.
abstract: Efficient tools for profiling DNA methylation in specific genes are essential for epigenetics and clinical diagnostics. Current DNA methylation profiling techniques have been limited by inconvenient implementation, requirements of specific reagents, and inferior accuracy in quantifying methylation degree. We develop a novel mass spectrometry method, target fragmentation assay (TFA), which enable to profile methylation in specific sequences. This method combines selective capture of DNA target from restricted cleavage of genomic DNA using magnetic separation with MS detection of the nonenzymatic hydrolysates of target DNA. This method is shown to be highly sensitive with a detection limit as low as 0.056 amol, allowing direct profiling of methylation using genome DNA without preamplification. Moreover, this method offers a unique advantage in accurately determining DNA methylation level. The clinical applicability was demonstrated by DNA methylation analysis using prostate tissue samples, implying the potential of this method as a useful tool for DNA methylation profiling in early detection of related diseases.
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Xiang-Cheng
authors.initials: XC
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Lan
authors.initials: L
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Ru-Qin
authors.initials: RQ
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University , Changsha 410082, P. R. China.
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D019175:DNA Methylation; D006801:Humans; D060526:Magnetic Fields; D008297:Male; D013058:Mass Spectrometry; D011467:Prostate / Q000378:metabolism
chemical_list: D004247:DNA