title: Three novel and one C31133T (Arg-338-->Stop) mutations of antihemophilic factor IX gene detected in Taiwan.
abstract: BACKGROUND
  Hemophilia-B is caused by mutation of the coagulation factor IX (F.IX) gene. The gene is about 34 kilobases in length and contains 8 exons ranging from 25 to 548 base pairs. There is no common mutation pattern of this gene in any population as there is in thalassemia. The hope is to localize the mutations in patients for use in making rapid familial surveys and for prenatal diagnoses.
  
  
  METHODS
  Diagnoses for a male patient with a newly-discovered mutation in Taiwan and three others with different novel mutations of F.IX gene were made from the results of prothrombin time (PT), activated partial thromboplastin time (APTT), assay of coagulation factors activity and factor IX antigen (F.IX:Ag) and also by assay of its inhibitor. The polymerase-chain-reaction (PCR)-based direct sequencing method was applied to evaluate all of the 8 exons and their flanking regions of F.IX gene for these patients.
  
  
  RESULTS
  The four male patients are all severe hemophilia-B. In the three patients with novel mutation of F.IX gene, two have a very low F.IX:Ag of less than 1% and one has 73%, but no one has any inhibitor. One of the four mutations is A to C transversion at nucleotide (nt) 10458 resulting in tyrosine (69) to serine change in epidermal growth factor (EGF) type B domain. Although the F.IX:Ag is present, the F.IX activity (F.IX:C) is markedly deficit owing to the change of beta-hydroxylase recognition site. Another two have mutations at the splicing junctions. They are G to C transversion at nt 118, i.e. the first base of IVS-1 splicing donor, and G to T transversion at nt 17668, the last base of IVS-4 splicing recipient. These mutations may cause splicing failure and subsequently result in deficiency of both F.IX activity and antigen. The other mutation that has never previously presented in this country is a C to T transition (arginine-338 to stop) at nt 31133 that results in early termination of translation.
  
  
  CONCLUSIONS
  The G to C mutation at nt 118 creates an Alu I restriction site and the C to T mutation at nt 31133 abolishes a Taq I site. Therefore, familial survey and rapid prenatal diagnosis for these patients by PCR-based direct sequencing method or by the restriction of enzyme cutting method pointing at the known mutation sites are certainly possible to achieve in our patients.
authors.affiliation: Department of Internal Medicine, Taichung Veterans General Hospital, Taiwan, R.O.C.
authors.forename: J H
authors.initials: JH
authors.lastname: Young
authors.forename: J C
authors.initials: JC
authors.lastname: Wang
authors.forename: Y C
authors.initials: YC
authors.lastname: Lin
mesh_terms: D001483:Base Sequence; D005164:Factor IX / Q000235:genetics*; D006467:Hemophilia A / Q000175:diagnosis*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D013624:Taiwan
chemical_list: D005164:Factor IX