title: Molecular analysis of the dystrophin gene in 407 Chinese patients with Duchenne/Becker muscular dystrophy by the combination of multiplex ligation-dependent probe amplification and Sanger sequencing.
abstract: BACKGROUND
  Progressive muscular dystrophy is a leading neuromuscular disorder without any effective treatments and a common genetic cause of mortality among teenagers. A challenge exists in the screening of subtle mutations in 79 exons and little is known about the genotype-phenotype correlation.
  
  
  METHODS
  Here we adopted multiplex ligation-dependent probe amplification and Sanger sequencing to detect the dystrophin gene in 407 patients and 76 mothers.
  
  
  RESULTS
  Sixty-three percent (257/407) of the patients harbored a deletion or duplication mutation, with a de novo mutation frequency of 39.5% in 76 affected patients, and approximately 43.7% of the deletions occurred from exon 45 to 52. To those patients suspected with single exon deletion, combined with Sanger sequencing, five subtle mutations were identified: c.8608C>T, c.2302C>T, c.7148dupT, c.10855C>T and c.2071-2093del AGGGAACAGATCCTGGTAAAGCA; the last three mutations were novel. Furthermore, after genotype-phenotype analysis, the severity of DMD/BMD was associated with the frame shift mutation but not with the deletion, the duplication or the number of deleted exons.
  
  
  CONCLUSIONS
  The majority of patients have a deletion/duplication mutation in the dystrophin gene, with a hot deletion mutation region from exon 45 to 52. Combined with Sanger sequencing, multiplex ligation-dependent probe amplification is capable of detecting part of subtle mutations.
authors.affiliation: Department of Neurology and Institute of Neurology, First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, China. wanjinchen75@yahoo.com.cn
authors.forename: Wan-Jin
authors.initials: WJ
authors.lastname: Chen
authors.forename: Qi-Fang
authors.initials: QF
authors.lastname: Lin
authors.forename: Qi-Jie
authors.initials: QJ
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: He
authors.forename: Xin-Yi
authors.initials: XY
authors.lastname: Liu
authors.forename: Min-Ting
authors.initials: MT
authors.lastname: Lin
authors.forename: Shen-Xing
authors.initials: SX
authors.lastname: Murong
authors.forename: Chia-Wei
authors.initials: CW
authors.lastname: Liou
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D002681:China; D016189:Dystrophin / Q000235:genetics*; D005260:Female; D005784:Gene Amplification; D056726:Genetic Association Studies; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D016189:Dystrophin