title: Genetic diagnosis of Duchenne and Becker muscular dystrophy using next-generation sequencing technology: comprehensive mutational search in a single platform.
abstract: BACKGROUND
  Duchenne muscular dystrophy or Becker muscular dystrophy might be a suitable candidate disease for application of next-generation sequencing in the genetic diagnosis because the complex mutational spectrum and the large size of the dystrophin gene require two or more analytical methods and have a high cost. The authors tested whether large deletions/duplications or small mutations, such as point mutations or short insertions/deletions of the dystrophin gene, could be predicted accurately in a single platform using next-generation sequencing technology.
  
  
  METHODS
  A custom solution-based target enrichment kit was designed to capture whole genomic regions of the dystrophin gene and other muscular-dystrophy-related genes. A multiplexing strategy, wherein four differently bar-coded samples were captured and sequenced together in a single lane of the Illumina Genome Analyser, was applied. The study subjects were 25
  
  
  METHODS
  16 with deficient dystrophin expression without a large deletion/duplication and 9 with a known large deletion/duplication.
  
  
  RESULTS
  Nearly 100% of the exonic region of the dystrophin gene was covered by at least eight reads with a mean read depth of 107. Pathogenic small mutations were identified in 15 of the 16 patients without a large deletion/duplication. Using these 16 patients as the standard, the authors' method accurately predicted the deleted or duplicated exons in the 9 patients with known mutations. Inclusion of non-coding regions and paired-end sequence analysis enabled accurate identification by increasing the read depth and providing information about the breakpoint junction.
  
  
  CONCLUSIONS
  The current method has an advantage for the genetic diagnosis of Duchenne muscular dystrophy and Becker muscular dystrophy wherein a comprehensive mutational search may be feasible using a single platform.
authors.affiliation: Department of Pediatrics, Seoul National University College of Medicine, Seoul National University Children's Hospital, Seoul, Korea.
authors.forename: Byung Chan
authors.initials: BC
authors.lastname: Lim
authors.forename: Seungbok
authors.initials: S
authors.lastname: Lee
authors.forename: Jong-Yeon
authors.initials: JY
authors.lastname: Shin
authors.forename: Jong-Il
authors.initials: JI
authors.lastname: Kim
authors.forename: Hee
authors.initials: H
authors.lastname: Hwang
authors.forename: Ki Joong
authors.initials: KJ
authors.lastname: Kim
authors.forename: Yong Seung
authors.initials: YS
authors.lastname: Hwang
authors.forename: Jeong-Sun
authors.initials: JS
authors.lastname: Seo
authors.forename: Jong Hee
authors.initials: JH
authors.lastname: Chae
mesh_terms: D000293:Adolescent; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis / Q000379:methods*; D016189:Dystrophin / Q000235:genetics*; D005091:Exons; D020440:Gene Duplication; D005838:Genotype; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis / Q000235:genetics; D010641:Phenotype; D017422:Sequence Analysis, DNA / Q000379:methods*; D017384:Sequence Deletion; D055815:Young Adult
chemical_list: D016189:Dystrophin