MLPA-based genotype–phenotype analysis in 1053 Chinese patients with DMD/BMD

BMC Medical Genetics

Cheng Zhang²  Xiao F Sun⁴  Jing Li²  Fei Chen⁴  Chang S Yu³  Yi X Zhan³  Yan Y Wang²  Shan W Feng¹  Ji Q Cao²  Ya Q Li²  Shao Y Li⁴  Juan Yang² 

[1] Institute of Population Research, Peking University, No.5 Yiheyuan Rd., Haidian District, Beijing City, 100080, People’s Republic of China;Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhong Shaner Road, Guangzhou City, 510080, People’s Republic of China;Guangzhou Kingmed Diagnostics Center, Guangzhou City, 510330, People’s Republic of China;Department of Reproductive Medicine, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou City, 510150, People’s Republic of China

关键词: MLPA;    DMD gene;    Becker muscular dystrophy;    Duchenne muscular dystrophy;   

Others  :  1177725
DOI  :  10.1186/1471-2350-14-29

 received in 2012-08-11, accepted in 2013-02-08,  发布年份 2013

【 摘 要 】

Background

Large-scale analysis of the transmission, mutation characteristics and the relationship between the reading frame and phenotype of the DMD gene has previously been performed in several countries, however, analogous studies have yet to be performed in Chinese populations.

Methods

Clinical data from 1053 Chinese patients with DMD/BMD were collected, and the DMD gene was tested by MLPA in all patients and 400 proband mothers. In 20 patients with negative MLPA, sequencing was also performed.

Results

We found that 27.50% of cases had a family medical history of DMD/BMD, and large rearrangements were identified in 70.56% of the probands, of which 59.35% and 11.21% were deletions or duplications, respectively. The carrier status of the mothers in the study was determined to be 50.75%, and it was established that the DMD mutation was inherited from the mother in 51.72% of the probands. Exons 45–54 and 3–22 were the most frequently deleted regions, and exons 3–11 and 21–37 were the most prevalently duplicated regions of the gene. Breakpoints mainly occurred in introns 43–55 for deletion mutations and in introns 2 and 7 for duplication mutations. No breakpoints were found at the 5^′ or 3^′ end of introns 31, 35, 36, 40, 65, 68, and 74–78 in any of the deletion or duplication mutations. The reading frame rule held true for 86.4% of the DMD patients and 74.55% of the BMD patients.

Conclusion

It is essential to increase physicians’ understanding of DMD/BMD, to promote scientific information, and to increase awareness in regards to genetic counseling and prenatal diagnosis in pedigrees with a family history of the disease, particularly in families with small DMD lesions in China. In addition, such a large-scale analysis will prove to be instructive for leading translational studies between basic science and clinical medicine.

【 授权许可】

   
2013 Yang et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Davies KE, Pearson PL, Harper PS, et al.: Linkage analysis of two cloned DNA sequences flanking the Duchenne muscular dystrophy locus on the short arm of the human X chromosome. Nucl Acids Res 1983, 11(8):2303-2312.
• [2]Kingston HM, Thomas NS, Pearson PL, et al.: Genetic linkage between Becker muscular dystrophy and a polymorphic DNA sequence on the short arm of the X chromosome. J Med Genet 1983, 20(4):255-258.
• [3]Hoffman EP, Brown RH Jr, Kunkel LM: Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 1987, 51(6):919-928.
• [4]Bushby KM, Thambyayah M, Gardner-Medwin D: Prevalence and incidence of Becker muscular dystrophy. Lancet 1991, 337(8748):1022-1024.
• [5]Emery AE: Population frequencies of inherited neuromuscular diseases: a world survey. Neuromuscul Disord 1991, 1(1):19-29.
• [6]Hoffman EP, Fischbeck KH, Brown RH, et al.: Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne’s or Becker’s muscular dystrophy. N Engl J Med 1988, 318(21):1363-1368.
• [7]Nicholson LV, Johnson MA, Gardner-Medwin D, et al.: Heterogeneity of dystrophin expression in patients with Duchenne and Becker muscular dystrophy. Acta Neuropathol 1990, 80(3):239-250.
• [8]Schouten JP, McElgunn CJ, Waaijer R, et al.: Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res 2002, 30(12):e57.
• [9]Roberts RG, Barby TF, Manners E, et al.: Direct detection of dystrophin gene rearrangements by analysis of dystrophin mRNA in peripheral blood lymphocytes. Am J Hum Genet 1991, 49(2):298-310.
• [10]den Dunnen JT, Antonarakis SE: Mutation nomenclature extensions and suggestions to describe complex mutations: a discussion. Hum Mutat 2000, 15(1):7-12.
• [11]Janssen B, Hartmann C, Scholz V, et al.: MLPA analysis for the detection of deletion, duplication and complex arrangements in the dystrophin gene: potential and pitfalls. Neurogenetics 2005, 6(1):29-35.
• [12]Gatta V, Scarciolla O, Gaspari AR, et al.: Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA). Hum Genet 2005, 117(1):92-98.
• [13]Tuffery-Giraud S, Béroud C, Leturcq F, et al.: Genotype–Phenotype Analysis in 2,405 Patients with a Dystrophinopathy Using the UMD–DMD Database: A Model of Nationwide Knowledgebase. Hum Mutat 2009, 30(6):934-945.
• [14]Hu XY, Ray PN, Worton RG: Mechanisms of tandem duplication in the Duchenne muscular dystrophy gene include both homologous and nonhomologous intrachromosomal recombination. EMBO J 1991, 10(9):2471-2477.
• [15]Zhang Z, Takeshima Y, Awano H, et al.: Tandem duplications of two separate fragments of the dystrophin gene in a patient with Duchenne muscular dystrophy. J Hum Genet 2008, 53(3):215-219.
• [16]White SJ, Aartsma-Rus A, Flanigan KM, et al.: Duplications in the DMD gene. Hum Mutat 2006, 27(9):938-945.
• [17]Gangopadhyay SB, Sherratt TG, Heckmatt JZ, et al.: Dystrophin in frameshift deletion patients with Becker muscular dystrophy. Am J Hum Genet 1992, 51(3):562-570.
• [18]Malhotra SB, Hart KA, Klamut HJ, et al.: Frame-shift deletions in patients with Duchenne and Becker muscular dystrophy. Science 1988, 242(4879):755-759.
• [19]Chelly J, Gilgenkrantz H, Lambert M, et al.: Effect of dystrophin gene deletions on mRNA levels and processing in Duchenne and Becker muscular dystrophies. Cell 1990, 63(6):1239-1248.
• [20]Gualandi F, Rimessi P, Trabanelli C, et al.: Intronic breakpoint definition and transcription analysis in DMD/BMD patients with deletion/duplication at the 5’ mutation hot spot of the dystrophin gene. Gene 2006, 370:26-33.
• [21]Winnard AV, Mendell JR, Prior TW, et al.: Frameshift deletions of exons 3–7 and revertant fibers in Duchenne muscular dystrophy: mechanisms of dystrophin production. Am J Hum Genet 1995, 56(1):158-166.
• [22]Yin H, Moulton HM, Seow Y, et al.: Cell-penetrating peptide-conjugated antisense oligonucleotides restore systemic muscle and cardiac dystrophin expression and function. Hum Mol Genet 2008, 17(24):3909-3918.
• [23]Cirak S, Arechavala-Gomeza V, Guglieri M, et al.: Exon skipping and dystrophin restoration in patients with Duchenne muscular dystrophy after systemic phosphorodiamidate morpholino oligomer treatment: an open-label, phase 2, dose-escalation study. Lancet 2011, 378(9791):595-605.