title: Mutations of the factor VIII gene in thai hemophilia A patients.
abstract: Hemophilia A is a common X-linked bleeding disorder caused by mutations in the coagulation factor VIII gene. The entire coding and essential sequences of the factor VIII gene were generated by a combination of genomic DNA amplification and long reverse transcription-polymerase chain reaction (long RT-PCR) using factor VIII transcripts prepared from lymphocytes. Mutations were then screened by non-radioactive single strand conformation polymorphism (SSCP) analysis and characterized by DNA sequencing. We have identified six potentially pathogenic mutations in the factor VIII gene in Thai hemophilia A patients, including two nonsense mutations (R-5X and R1966X), three missense mutations (D542Y, G1850V, and G2325C), and a 4-bp insertion (ACTA) at codon 2245. Three of these mutations (D542Y, G2325C, and 4-bp insertion) have never been previously reported, and the ins2245 is the first example of such insertion probably causing factor VIII elongation. R1966X, D542Y, G1850V, and 4-bp insertion were associated with a severe hemophiliac phenotype whereas R-5X and G2325C were observed in moderately affected patients. Mutations in the factor VIII gene in Thai hemophilia A patients are likely to be heterogeneous. This study represents the first attempt to further the understanding of the molecular basis of hemophilia A in Thai.
authors.affiliation: Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, Nakhon Pathom.
authors.forename: V
authors.initials: V
authors.lastname: Akkarapatumwong
authors.forename: S
authors.initials: S
authors.lastname: Oranwiroon
authors.forename: P
authors.initials: P
authors.lastname: Pung-amritt
authors.forename: A
authors.initials: A
authors.lastname: Treesucon
authors.forename: P
authors.initials: P
authors.lastname: Thanootarakul
authors.forename: G
authors.initials: G
authors.lastname: Veerakul
authors.forename: C
authors.initials: C
authors.lastname: Mahasandana
authors.forename: S
authors.initials: S
authors.lastname: Panyim
authors.forename: P
authors.initials: P
authors.lastname: Yenchitsomanus
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002570:Ceruloplasmin / Q000235:genetics; D005165:Factor V / Q000235:genetics; D005169:Factor VIII / Q000235:genetics*; D005260:Female; D006467:Hemophilia A / Q000235:genetics*; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013552:Swine; D013785:Thailand
chemical_list: D005165:Factor V; D005169:Factor VIII; D002570:Ceruloplasmin