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

Affiliations
  • 1Department of Pediatrics, Pusan Paik Hospital, Inje University College of Medicine, Busan, Korea.
  • 2Department of Pediatrics, Eulji General Hospital, Seoul, Korea.
  • 3Department of Pediatrics, Samsung Changwon Hospital, Sungkyunkwan University School of Medicine, Changwon, Korea.
  • 4Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
  • 5Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. jhnr.lee@samsung.com

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.


Cited by  2 articles

Multiplex Ligation-Dependent Probe Amplification in X-linked Recessive Muscular Dystrophy in Korean Subjects
Mi Ri Suh, Kyung-A Lee, Eun Young Kim, Jiho Jung, Won Ah Choi, Seong-Woong Kang
Yonsei Med J. 2017;58(3):613-618.    doi: 10.3349/ymj.2017.58.3.613.

Duchenne Muscular Dystrophy and Becker Muscular Dystrophy Confirmed by Multiplex Ligation-Dependent Probe Amplification: Genotype-Phenotype Correlation in a Large Cohort
Seena Vengalil, Veeramani Preethish-Kumar, Kiran Polavarapu, Manjunath Mahadevappa, Deepha Sekar, Meera Purushottam, Priya Treesa Thomas, Saraswathi Nashi, Atchayaram Nalini
J Clin Neurol. 2017;13(1):91-97.    doi: 10.3988/jcn.2017.13.1.91.


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