title: [Molecular characterization of genetic defects in hemophilia in Shanghai].
abstract: OBJECTIVE
  To detect gene mutations in hemophilia A in Shanghai.
  
  
  METHODS
  PCR, denaturing gradient gel electrophoresis (DGGE) and DNA sequencing were used. Fifty Chinese cases of hemophilia A without intron 22 inversion, including 24 severe, 9 moderate and 17 mild cases, were screened. Genomic DNA was amplified using GC-clamped primers covering all the exons and flanking intron regions, excluding the middle portion of extron 14 encoding the B domain, but including the thrombin cleavage sites at AA740 and 1689. The amplified GC-clamped PCR fragments were then electrophoresed on DGGE. The abnormal bands were sequenced.
  
  
  RESULTS
  Eleven different mutations were identified, including 5 nonsense mutations, 5 missense mutations and one small deletion. Among them, 3 mutations, 466Lys (AAG)-Thr (ACG), 719Tyr (TAC)-Stop (TAG) and 312Ile (ATC)-xxC have not been reported before.
  
  
  CONCLUSIONS
  Apart from intron 22 inversion, most gene mutations in hemophilia A were point mutations resulted from single base substitution. Generally the genetic defects correspond to the clinical manifestations.
authors.affiliation: Shanghai Institute of Hematology, Rui jin Hospital, Shanghai Second Medical University, Shanghai 200025.
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.forename: W
authors.initials: W
authors.lastname: Huang
authors.forename: W
authors.initials: W
authors.lastname: Cao
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: B
authors.initials: B
authors.lastname: Qu
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Shao
authors.forename: Z
authors.initials: Z
authors.lastname: Wang
authors.forename: L
authors.initials: L
authors.lastname: Chen
authors.forename: W
authors.initials: W
authors.lastname: Huang
mesh_terms: D044466:Asian People / Q000235:genetics*; D002681:China; D006467:Hemophilia A / Q000235:genetics*; D006801:Humans; D007438:Introns / Q000235:genetics; D009154:Mutation