Methyl-CpG-Binding PCR of Bloodspots for Confirmation of Fragile X Syndrome in Males

[摘要] This study demonstrates that methyl-CpG-binding PCR (MB-PCR) is a rapid andsimple method for detecting fragile X syndrome (FXS) in males, which isperformed by verifying the methylation status of theFMR1promoter inbloodspots. Proteins containing methyl-CpG-binding (MB) domains can befreeze-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 unmethylatedalleles up to 320 folds. We examined bloodspots from 100 males, including 24with FXS, in a blinded manner. The results revealed that the ability of MB-PCR todetectFMR1promoter methylation was the same as that of Southern blothybridization. Since individuals with 2 or more X chromosomes generally havemethylatedFMR1alleles, MB-PCR cannot be used to detect FXS in females.

[学科分类] 基础医学