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Journal of Biomedicine and Biotechnology
Volume 2009, Article ID 643692, 8 pages
Methodology Report

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

1Department of Pathology, Chi Mei Medical Center, Tainan, Taiwan
2Department of Pediatrics, Chi Mei Medical Center, Tainan, Taiwan
3The Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Kaohsiung, Taiwan
4Department of Pediatrics, Buddhist Tzu Chi General Hospital, Taipei Branch, Taipei, Taiwan

Received 28 February 2009; Revised 30 June 2009; Accepted 10 August 2009

Academic Editor: Wolfgang Schulz

Copyright © 2009 Ching-Cherng Tzeng et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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.