Browse > Article

Genetic Diagnosis of Beckwith Wiedemann Syndrome using Methylation Specific PCR-RFLP Method  

Kim, Gu-Hwan (Medical Genetics Center, Asan Medical Center Children's Hospital)
Lee, Jin-Joo (Medical Genetics Center, Asan Medical Center Children's Hospital)
Choi, Seung-Hoon (Medical Genetics Center, Asan Medical Center Children's Hospital)
Lee, Joo-Yeon (Medical Genetics Center, Asan Medical Center Children's Hospital)
Lee, Beom-Hee (Medical Genetics Center, Asan Medical Center Children's Hospital)
Yoo, Han-Wook (Medical Genetics Center, Asan Medical Center Children's Hospital)
Publication Information
Journal of Genetic Medicine / v.7, no.2, 2010 , pp. 133-137 More about this Journal
Abstract
Purpose: Beckwith-Wiedemann syndrome (BWS) is an overgrowth malformation syndrome caused by a methylation abnormality at chromosome 11p15, consisting of two imprinting centers, BWSIC1 (IGF2, H19) and BWSIC2 (LIT1, KvDMR). This study evaluated the applicability of a methylation-specific (MS) PCR RFLP method for the genetic diagnosis of BWS. Materials and Methods: A total of 12 patients were recruited based on clinical findings. Karyotyping was performed using peripheral blood leukocytes, and genomic DNA was treated with bisulfate and amplified using methylation-specific primers. RFLP was conducted with restriction enzymes in differentially methylated regions of LIT1, H19, and IGF2. Results: The 12 BWS patients had normal karyotypes. Abnormal methylation patterns in the BWSIC2 (LIT1) region were identified in seven patients (58.3%) using the MS-PCR RFLP method. Conclusions: The MS-PCR RFLP method is a simple, economical genetic test. It detected genetic abnormalities in 50-60% of BWS patients, suggesting that it can be used as a screening test. A more precise method is required, however, to enhance the detection rate of genetic abnormalities, especially in BWSIC1 region.
Keywords
Beckwith-Wiedemann syndrome; Methylation; Genetic testing; RFLP; BWSIC1; BWSIC2; LIT1;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Ulaner GA, Vu TH, Li T, Hu JF, Yao XM, Yang Y, et al. Loss of imprinting of IGF2 and H19 in oseteosarcoma is accompanied by reciprocal methylation changes of CTCF-binding site. Hum Mol Genet 2003;12:535-49.   DOI
2 Kim GH, Lee BH, Yoo HW. MLPA applications in genetic testing. J Genet Med 2009;6:146-54.
3 Shuman C, Smith AC, Weksberg R. Beckwith-Wiedemann syndrome. 2008. May. GenReviews PMID20301568. Available from: http://www.genetests.org
4 Li M, Squire J, Shuman C, Fei YL, Atkin J, Pauli R, et al. Imprinting status of 11p15 genes in Beckwith-Wiedemann syndrome patients with CDKN1C mutations. Genomics 2001;74:370-6.   DOI   ScienceOn
5 Weksberg R, Shuman C, Smith AC. Beckwith-Wiedemann syndrome. Am J Med Genet C Semin Med Genet 2005;137:12-23.
6 Frommer M, McDonald LE, Millar DS, Collis CM, Watt F, Grigg GW, et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. Proc Natl Acad Sci 1992;89:1827-31.   DOI   ScienceOn
7 Wiedemann HR. Familial malformation complex with umbilical hernia and macroglossia: "A new syndrome?" J Genet Hum 1964;13:223-32.
8 Reik W, Maher ER. Imprinting in clusters: lessons from Beckwith-Wiedemann syndrome. Trends Genet 1997;13:330-4.   DOI   ScienceOn
9 Bliek J, Maas SM, Ruijter JM, Hennekam RC, Alders M, Westerveld A, et al. Increased tumour risk for BWS patients correlates with aberrant H19 and not KCNQ1OT1 methylation: occurrence of KCNQ1OT1 hypomethylation in familial cases of BWS. Hum Mol Genet 2001;10:467-76.   DOI
10 Scott RH, Douglas J, Baskcomb L, Nygren AO, Birch JM, Cole TR, et al. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) robustly detects and distinguishes 11p15 abnormalities associated with overgrowth and growth retardation. J Med Genet 2008;45:106-13.