J Korean Med Sci.  2012 Mar;27(3):274-280. 10.3346/jkms.2012.27.3.274.

Genetic Analysis of Dystrophin Gene for Affected Male and Female Carriers with Duchenne/Becker Muscular Dystrophy in Korea

• Lee BL
• Nam SH
• Lee JH
• Ki CS
• Lee M
• Lee J

• KMID: 1792998
• DOI: http://doi.org/10.3346/jkms.2012.27.3.274

Abstract

Duchenne and Becker muscular dystrophy (DMD/BMD) are X-linked recessive disorders caused by mutation in dystrophin gene. We analyzed the results of a genetic test in 29 DMD/BMD patients, their six female relatives, and two myopathic female patients in Korea. As the methods developed, we applied different procedures for dystrophin gene analysis; initially, multiplex polymerase chain reaction was used, followed by multiplex ligation-dependent probe amplification (MLPA). Additionally, we used direct DNA sequencing for some patients who had negative results using the above methods. The overall mutation detection rate was 72.4% (21/29) in DMD/BMD patients, identifying deletions in 58.6% (17/29). Most of the deletions were confined to the central hot spot region between exons 44 and 55 (52.9%, 7/19). The percentage of deletions and duplications revealed by MLPA was 45.5% (5/11) and 27.2% (3/11), respectively. Using the MLPA method, we detected mutations confirming their carrier status in all female relatives and symptomatic female patients. In one patient in whom MLPA revealed a single exon deletion of the dystrophin gene, subsequent DNA sequencing analysis identified a novel nonsense mutation (c.4558G > T; Gln1520X). The MLPA assay is a useful quantitative method for detecting mutation in asymptomatic or symptomatic carriers as well as DMD/BMD patients.

Keyword

Gene Amplification; Duchenne/Becker Muscular Dystrophy; Deletion; Duplication

MeSH Terms

Adolescent
Adult
Child
Child, Preschool
DNA Mutational Analysis
Dystrophin/*genetics
Exons
Female
Heterozygote
Humans
Infant
Ligase Chain Reaction
Male
Multiplex Polymerase Chain Reaction
Muscular Dystrophy, Duchenne/*genetics
Mutagenesis, Insertional
Republic of Korea
Sequence Analysis, DNA
Sequence Deletion

Figure

• 

  Fig. 1 Duplications of exons 3-7 of the dystrophin gene in one affected patient (case 23). (A) Electrophoregrams after MLPA in a control patient, (B) Electrophoregrams after MLPA in case 23.

• 

  Fig. 2 Deletions of exons 18-44 of the dystrophin gene in one affected patient (case 21) and his mother (case 30). These electrophoregrams show the MLPA sample after hybridization with SALSA probe mix P034 (dystrophin gene exons 1-10, 21-30, 41-50, and 61-70). The deletions of exons 18-20 and 31-40 were detected after probe hybridization with P035 (dystrophin gene exons 11-20, 31-40, 51-60, and 71-79), which is not shown. (A) Control, (B) Female carrier, (C) Case 21.

• 

  Fig. 3 The results of single exon PCR (A) and electrophoregrams after MLPA analysis (B) of exon 33 in the dystrophin gene of case 20. (A) Single exon PCR result. Lane 1 is from a control sample, and the band of lane 2 corresponds with exon 33 of case 20. (B) The arrow indicates absence of a peak corresponding to exon 33 in MLPA analysis.

• 

  Fig. 4 Direct DNA sequencing of exon 33. The arrow indicates the hemizygous G to T transversion at nucleotide 4558 in exon 33 (c.4558G > T;Glu1520X) of the dystrophin gene of case 20. (A) Control, (B) Case 20.

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