title: A quantitative DNA methylation assay using mismatch hybridization and chemiluminescence.
abstract: OBJECTIVE
  To develop a quantitative method for methylation analysis of the p16 gene based on mismatch hybridization and chemiluminescence.
  
  
  METHODS
  Genomic DNA was modified by sodium bisulfite to convert all unmethylated but not methylated cytosines to uracil, and subsequently a pair of primer having no CpG sites was designed for amplification target DNA containing methylated or unmethylated CpG sites. The PCR product spanning CpG sites were hybridized with two oligonucleotide probes which perfectly matched the methylated and unmethylated CpG sequences respectively, and the hybrids were detected by chemiluminescent method. The percentage of methylated target sequences could be estimated by calculating the ratio of signals obtained with two probes.
  
  
  RESULTS
  The percentage of methylation of artificial mixtures DNA showed a linear relation. There was a negative correlation between the methyaltion index with p16 transcriptional mRNA of p16 gene in tumor cell lines.
  
  
  CONCLUSIONS
  Compared with existing methods, this assay is nonisotopic, rapid, simple, and can be widely applied to the study of DNA methylation.
authors.affiliation: Institute of Environmental Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors.forename: Qun-Feng
authors.initials: QF
authors.lastname: Yao
authors.forename: Xin-Jiang
authors.initials: XJ
authors.lastname: Kang
authors.forename: Qiao-Ling
authors.initials: QL
authors.lastname: Hao
authors.forename: Yi-Kai
authors.initials: YK
authors.lastname: Zhou
mesh_terms: D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D019942:Genes, p16; D006801:Humans; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013447:Sulfites
chemical_list: D013447:Sulfites; C009279:sodium bisulfite