Abstract：The purpose of this study is to analyze the family's clinical data of 22 children who were given an intended clinical diagnosis of Duchenne muscular dystrophy (DMD), and to explore the clinical value of next-generation sequencing (NGS) in the molecular diagnosis of DMD. The probands were simultaneously tested by NGS for a gene panel associated with hereditary neuromuscular disease and multiplex ligation-dependent probe amplifcation (MLPA) for the Dystrophin gene. The exon deletion/repetition mutations of the Dystrophin gene determined by both methods were compared and the point mutations of the Dystrophin gene were verifed by Sanger sequencing. Dystrophin gene mutations were found in all the 22 probands, including 14 exon deletion/repetition mutations and 8 point mutations/minor variations. The results of MLPA detection were consistent with those of NGS. The results of Sanger sequencing showed that the point mutations and minor variations determined by NGS were correct. One missense mutation (c.6290G>T), 1 nonsense mutation (c.3487C>T) and 4 minor deletion-induced frameshift mutations (c.1208delG, c.7497_7506delGGTGGGTGAC, c.9421_9422delAA and c.8910_8913delTCTC) had not been reported in the Human Gene Mutation Database, and thus were considered as novel mutations of the Dystrophin gene. The results of this study showed that NGS can detect variations in the Dystrophin gene, including exon deletion/repetition, point mutation, minor deletion and intron mutation. Therefore, NGS is of certain clinical value in the molecular diagnosis of DMD and is worthy of recommendation.
TIAN Pei-Chao,WANG Yue,SHI Dan-Dan et al. Application of next-generation sequencing in the molecular diagnosis of Duchenne muscular dystrophy[J]. CJCP, 2019, 21(3): 244-248.
Birnkrant DJ, Bushby K, Bann CM, et al. Diagnosis and management of Duchenne muscular dystrophy, part 1:diagnosis, and neuromuscular, rehabilitation, endocrine, and gastrointestinal and nutritional management[J]. Lancet Neurol, 2018, 17(3):251-267.
[2]
Freund AA, Scola RH, Arndt RC, et al. Duchenne and Becker muscular dystrophy:a molecular and immunohistochemical approach[J]. Arq Neuropsiquiatr, 2007, 65(1):73-76.
Aartsma-Rus A, Van Deutekom JC, Fokkema IF, et al. Entries in the Leiden Duchenne muscular dystrophy mutation database:an overview of mutation types and paradoxical cases that confrm the reading-frame rule[J]. Muscle Nerve, 2006, 34(2):135-144.
Lim BC, Lee S, Shin JY, et al. Genetic diagnosis of Duchenne and Becker muscular dystrophy using next-generation sequencing technology:comprehensive mutational search in a single platform[J]. J Med Genet, 2011, 48(11):731-736.
McKenna A, Hanna M, Banks E, et al. The Genome Analysis Toolkit:a MapReduce framework for analyzing next-generation DNA sequencing data[J]. Genome Res, 2010, 20(9):1297-1303.
[9]
DePristo MA, Banks E, Poplin R, et al. A framework for variation discovery and genotyping using next-generation DNA sequencing data[J]. Nat Genet, 2011, 43(5):491-498.
[10]
Krumm N, Sudmant PH, Ko A, et al. Copy number variation detection and genotyping from exome sequence data[J]. Genome Res, 2012, 22(8):1525-1532.
[11]
Fowler A, Mahamdallie S, Ruark E, et al. Accurate clinical detection of exon copy number variants in a targeted NGS panel using DECoN[J]. Wellcome Open Res, 2016, 1:20.
Takeshima Y, Yagi M, Okizuka Y, et al. Mutation spectrum of the dystrophin gene in 442 Duchenne/Becker muscular dystrophy cases from one Japanese referral center[J]. J Hum Genet, 2010, 55(6):379-388.
[17]
Juan-Mateu J, Gonzalez-Quereda L, Rodriguez MJ, et al. DMD mutations in 576 dystrophinopathy families:a step forward in genotype-phenotype correlations[J]. PLoS One, 2015, 10(8):e0135189.
