title: [Mutation screening of the F VIII gene in 10 hemophilia A families].
abstract: OBJECTIVE
  To identify the F VIII gene mutations of patients and suspected female carriers in 10 Hemophilia A (HA) families, and to guide the prenatal diagnosis.
  
  
  METHODS
  PCR, denaturinghigh performance liquid chromatogramphy (DHPLC) and DNA sequencing technologies were applied to screen the F VIII gene of 8 HA patients and 12 suspected female carriers in the 10 families. Linkage analysis was performed by using St 14(DXS 52), intron 13 (CA)n and EX18/Bcl I of the F VIII gene in the HA families. In prenatal diagnosis, we screened the same mutation found in the patients. PCR-restriction fragment length polymorphism was applied to detect the new missense mutations of F VIII gene in 100 unrelated healthy individuals to exclude the possibility of polymorphism.
  
  
  RESULTS
  Five missense mutations, 3 frameshift mutations, 2 nonsense mutations and 2 single nucleotide polymorphism (SNP) were identified in 10 the HA families. Among them, c.878A to G, c.1015A to G, c.6870G to T, c.1282delA, c.3072_3073insT, c.4880_4881insA and c.5000G to A were novel mutations or polymorphism. No missense mutations c.878A G, c.1015A to G and c.6870G to T, were found in the 100 healthy unrelated controls. (2) Nine suspected female carriers were confirmed at the gene level. (3) X risk chromosome could be determined to in 4 HA families by genetic linkage analysis. (4) Among the four fetuses for prenatal diagnosis, 2 were normal, 1 was carrier and the remaining 1 was a patient.
  
  
  CONCLUSIONS
  Six novel mutations, i.e., c.878A to G, c.1015A to G, c.6870G to T, c.1282delA, c.3072_3073insT and c.4880_4881insA, were identified in this study. PCR, DHPLC and DNA sequencing could be used to screen the gene mutations of HA patients, to carry out carrier detection and prenatal diagnosis of HA families efficiently, by combining with restriction endonuclease analysis and genetic linkage analysis.
authors.affiliation: Institute of Reproductive and Stem Cell Engineering of Central South University, Changsha, Hunan,410008 P.R. China. Lugxdirector @yahoo.com.cn
authors.forename: Wen
authors.initials: W
authors.lastname: LI
authors.forename: Xiao
authors.initials: X
authors.lastname: HU
authors.forename: Bo-di
authors.initials: BD
authors.lastname: GAO
authors.forename: Lu-yun
authors.initials: LY
authors.lastname: LI
authors.forename: Yi
authors.initials: Y
authors.lastname: LIAO
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: TANG
authors.forename: Wei-lin
authors.initials: WL
authors.lastname: TANG
authors.forename: Guang-xiu
authors.initials: GX
authors.lastname: Lu
mesh_terms: D041321:Chromosomes, Human, X; D004252:DNA Mutational Analysis / Q000379:methods; D004262:DNA Restriction Enzymes / Q000235:genetics; D005169:Factor VIII / Q000235:genetics*; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006467:Hemophilia A / Q000175:diagnosis / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D008297:Male; D009154:Mutation; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D011296:Prenatal Diagnosis / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D005169:Factor VIII; D004262:DNA Restriction Enzymes