title: Methyl-CpG-binding PCR of bloodspots for confirmation of fragile X syndrome in males.
abstract: This study demonstrates that methyl-CpG-binding PCR (MB-PCR) is a rapid and simple method for detecting fragile X syndrome (FXS) in males, which is performed by verifying the methylation status of the FMR1 promoter in bloodspots. Proteins containing methyl-CpG-binding (MB) domains can be freeze-stored and used as stocks, and the entire test requires only a few hours. The minimum amount of DNA required for the test is 0.5 ng. At this amount, detection sensitivity is not hampered, even mixing with excess unmethylated alleles up to 320 folds. We examined bloodspots from 100 males, including 24 with FXS, in a blinded manner. The results revealed that the ability of MB-PCR to detect FMR1 promoter methylation was the same as that of Southern blot hybridization. Since individuals with 2 or more X chromosomes generally have methylated FMR1 alleles, MB-PCR cannot be used to detect FXS in females.
authors.affiliation: Department of Pathology, Chi Mei Medical Center, Tainan, Taiwan. tzeng-tainan@yahoo.com.tw
authors.forename: Ching-Cherng
authors.initials: CC
authors.lastname: Tzeng
authors.forename: Chiou-Ping
authors.initials: CP
authors.lastname: Liou
authors.forename: Chien-Feng
authors.initials: CF
authors.lastname: Li
authors.forename: Ming-Chi
authors.initials: MC
authors.lastname: Lai
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Tsai
authors.forename: Wei-Chen
authors.initials: WC
authors.lastname: Cho
authors.forename: Hui-Ting
authors.initials: HT
authors.lastname: Chang
mesh_terms: D001800:Blood Specimen Collection / Q000379:methods*; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000097:blood* / Q000235:genetics*; D005600:Fragile X Syndrome / Q000097:blood* / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human