title: [Detecting testis- and epididymis-specific methylated promoters in human cell-free seminal DNA by MeDIP-qPCR].
abstract: OBJECTIVE
  To establish a method of methyl-DNA immunoprecipitation (MeDIP)-real time quantitative PCR (qPCR) for detecting the promoter methylation level in cell-free seminal DNA (cfsDNA).
  
  
  METHODS
  We obtained cfsDNA samples from 6 normozoospermia men (the NZ group) and 6 post-vasectomy patients (the PV group), and mixed the samples from different individuals of each group, respectively. Then we made DNA fragments by ultrasonication, separated the methylated DNA fragments by MeDIP, and determined the methylation level of the promoters in cfsDNA by qPCR.
  
  
  RESULTS
  The methylation levels of the promoters PRAME, PEG10, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4, DAZ1 and CLPB were 14.93, 2.64, 0.69, 2.66, 17.50, 21.10, 5.98, 2.28, 13.50 and 3.86%, respectively, in the NZ group, obviously lower than 121.25, 73.62, 16.25, 42.90, 76.74, 112.40, 59.79, 25.85, 91.90 and 64.53% in the PV group. The results of MeDIP-qPCR for the methylation of PRAME, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4 and DAZ1 were coincident with the results of genome-wide promoter methylation microarray.
  
  
  CONCLUSIONS
  MeDIP-qPCR can quantitatively measure the promoter methylation level in cfsDNA, and effectively determine the testis- and epididymis-specific methylated promoters in human semen.
authors: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors: Huang-Tao
authors: HT
authors: Guan
authors: Family Planning Service Station of Qichun County, Huanggang, Hubei 435300, China.
authors: Jin-Ming
authors: JM
authors: Lü
authors: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors: Chun-Lin
authors: CL
authors: Wu
authors: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors: Hong-Gang
authors: HG
authors: Li
authors: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors: Chang-Hong
authors: CH
authors: Zhu
authors: Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors: Cheng-Liang
authors: CL
authors: Xiong
mesh_terms: D000328:Adult; D004247:DNA / Q000737:chemistry; D019175:DNA Methylation; D004822:Epididymis / Q000378:metabolism*; D044127:Epigenesis, Genetic; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012661:Semen / Q000737:chemistry*; D013737:Testis / Q000378:metabolism*
chemical_list: D004247:DNA