Multiplex Ligation-dependent Probe Amplification (MLPA) 방법에 의한 디스트로핀 유전자 돌연변이 분자학적 진단의 유용성
Clinical Usefulness of Molecular Diagnosis in Dystrophin Gene Mutations Using the Multiplex Ligation-dependent Probe Amplification (MLPA) Method
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Background: Duchenne/Becker muscular dystrophy (DMD/BMD), which is the most common X-linked muscular dystrophy, is caused by mutations in the dystrophin gene. These mutations comprise deletions in approximately 55~65% of patients, duplications in 5~10%, and point mutations or small insertion/deletions in the remainder. Unfortunately, current diagnostic assays for dystrophin do not accurately detect duplication mutations or female carriers. In this study we employed multiplex ligation-dependent probe amplification (MLPA) analysis to detect deletions or duplications of the dystrophin gene in patients with DMD/BMD, and in potential female carriers. Methods: A total of 41 subjects was recruited for this study, comprising 35 male DMD/BMD patients, 1 female patient with Turner syndrome, and 5 females with a family history of DMD/BMD. The MLPA method was employed to determine the copy number of each of the 79 exons of the dystrophin gene in the 41 subjects. Results: MLPA analysis for dystrophin was informative in 71.4% (25/35) of patients with DMD/BMD patients, identifying deletions in 60.0% (21/35) and duplications in 11.4% (4/35). MLPA analysis showed the presence of a deletion of the DMD gene in one female patient with Turner syndrome. Of the five female patients with a family history of DMD/BMD, this assay revealed exon deletion in one and duplications in one. Conclusions: The reported findings reveal that the MLPA method is a powerful tool for detecting duplications and female carriers, as well as DMD gene deletions. MLPA should be considered the method of choice for an initial genetic analysis of DMD/BMD patients.
Abstract
Background: Duchenne/Becker muscular dystrophy (DMD/BMD), which is the most common X-linked muscular dystrophy, is caused by mutations in the dystrophin gene. These mutations comprise deletions in approximately 55~65% of patients, duplications in 5~10%, and point mutations or small insertion/deletions in the remainder. Unfortunately, current diagnostic assays for dystrophin do not accurately detect duplication mutations or female carriers. In this study we employed multiplex ligation-dependent probe amplification (MLPA) analysis to detect deletions or duplications of the dystrophin gene in patients with DMD/BMD, and in potential female carriers. Methods: A total of 41 subjects was recruited for this study, comprising 35 male DMD/BMD patients, 1 female patient with Turner syndrome, and 5 females with a family history of DMD/BMD. The MLPA method was employed to determine the copy number of each of the 79 exons of the dystrophin gene in the 41 subjects. Results: MLPA analysis for dystrophin was informative in 71.4% (25/35) of patients with DMD/BMD patients, identifying deletions in 60.0% (21/35) and duplications in 11.4% (4/35). MLPA analysis showed the presence of a deletion of the DMD gene in one female patient with Turner syndrome. Of the five female patients with a family history of DMD/BMD, this assay revealed exon deletion in one and duplications in one. Conclusions: The reported findings reveal that the MLPA method is a powerful tool for detecting duplications and female carriers, as well as DMD gene deletions. MLPA should be considered the method of choice for an initial genetic analysis of DMD/BMD patients.
참고문헌 (27)
- Emery AE, "Population frequencies of inherited neuromuscular diseases--a world survey", Neuromuscul Disord, 1, pp.19~29 (1991)
- Prior TW, "Experience and strategy for the molecular testing of Duchenne muscular dystrophy", J Mol Diagn, 7, pp.317~326 (2005)
- Ervasti JM, "Membrane organization of the dystrophinglycoprotein complex", Cell, 66, pp.1121~1131 (1991)
- Ervasti JM, "A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin", J Cell Biol, 122, pp.809~823 (1993)
- Forrest SM, "Further studies of gene deletions that cause Duchenne and Becker muscular dystrophies", Genomics, 2, pp.109~114 (1988)
- Koenig M, "Complete cloning of the Duchenne muscular dystrophy (DMD)cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals", Cell, 50, pp.509~517 (1987)
- Monaco AP, "An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus", Genomics, 2, pp.90~95 (1988)
- Den Dunnen JT, "Topography of the Duchenne muscular dystrophy (DMD) gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications", Am J Hum Genet, 45, pp.835~847 (1989)
- Hu XY, "Duplicational mutation at the Duchenne muscular dystrophy locus: its frequency, distribution, origin, and phenotypegenotype correlation", Am J Hum Genet, 46, pp.682~695 (1990)
- Roberts RG, "Point mutations in the dystrophin gene", Proc Natl Acad Sci U S A, 89, pp.2331~2335 (1992)
- Abbs S, "A convenient multiplex PCR system for the detection of dystrophin gene deletions: a comparative analysis with cDNA hybridisation shows mistypings by both methods", J Med Genet, 28, pp.304~311 (1991)
- Beggs AH, "Detection of 98% of DMD", BMD gene deletions by polymerase chain reaction, 1990, pp.86, 45~48
- 나상준, "Dystrophinopathy 환자의 임상적, 면역조직화학적 및 유전학적 분석", 대한신경과학회지, 22(5), pp.508~515 (2004), 대한신경과학회
- Yamagishi H, "Identification of carriers of Duchenne", Becker muscular dystrophy by a novel method based on detection of junction fragments in the dystrophin gene, 1996, pp.33, 1027~1031
- Yau SC, "Accurate diagnosis of carriers of deletions and duplications in Duchenne", Becker muscular dystrophy by fluorescent dosage analysis, 1996, pp.33, 550~558
- Xiao Y, "Screening for DMD", BMD deletion carriers by fluorescence in situ hybridization, 2003, pp.7, 195~201
- Schwartz M, "Improved molecular diagnosis of dystrophin gene mutations using the multiplex ligation-dependent probe amplification method", Genet Test, 8, pp.361~367 (2004)
- Sellner LN, "MLPA and MAPH: new techniques for detection of gene deletions", Hum Mutat, 23, pp.413~419 (2004)
- 박윤희, "듀센형근이영양증 유전자의 결실 돌연변이 검출을 위한 Dual Priming Oligonucleotide 다중 PCR법의 평가", The Korean Journal of Laboratory Medicine, 28(5), pp.386~391 (2008), 대한진단검사의학회
- Schouten JP, "Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification", Nucleic Acids Res, 30, pp.57~ (2002)
- Hwa HL, "Multiplex ligation-dependent probe amplification identification of deletions and duplications of the Duchenne muscular dystrophy gene in Taiwanese subjects", J Formos Med Assoc, 106, pp.339~346 (2007)
- Imoto N, "Topographic pattern of the rearrangement of the dystrophin gene in Japanese Duchenne muscular dystrophy", Hum Genet, 92, pp.533~536 (1993)
- Onengut S, "Deletion pattern in the dystrophin gene in Turks and a comparison with Europeans and Indians", Ann Hum Genet, 64, pp.33~40 (2000)
- Hu XY, "Evidence for mutation by unequal sister chromatid exchange in the Duchenne muscular dystrophy gene", Am J Hum Genet, 44, pp.855~863 (1989)
- Robinson WP, "The mechanisms involved in formation of deletions and duplications of 15q11-q13", J Med Genet, 35, pp.130~136 (1998)
- Potocki L, "Molecular mechanism for duplication 17p11.2- the homologous recombination reciprocal of the Smith-Magenis microdeletion", Nat Genet, 24, pp.84~87 (2000)
- Lee KA, "Becker muscular dystrophy with r(X) carrying an out-of-frame DMD deletion", Pediatr Neurol, 39, pp.129~132 (2008)