Methyl-CpG-binding PCR of bloodspots for confirmation of fragile X syndrome in males.
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.
Tzeng
Ching-Cherng
CC
Department of Pathology, Chi Mei Medical Center, Tainan, Taiwan. tzeng-tainan@yahoo.com.tw
Liou
Chiou-Ping
CP
Li
Chien-Feng
CF
Lai
Ming-Chi
MC
Tsai
Li-Ping
LP
Cho
Wei-Chen
WC
Chang
Hui-Ting
HT
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
D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human