Original Paper

Cell Biochemistry and Biophysics

, Volume 65, Issue 3, pp 463-472

A Strategy for the Molecular Diagnosis in Hemophilia A in Chinese Population

  • Zhihui HeAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Juan ChenAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Shiyan XuAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Shufen ChenAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Xiao XiaoAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Hongyi LiAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Yibin GuoAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University
  • , Weiying JiangAffiliated withDepartment of Medical Genetics, Medical School and Key Laboratory of Tropic Disease Control, Ministry Education, Sun Yat-sen University Email author 

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Abstract

Hemophilia A is an x-linked recessive inherited bleeding disorder. So far, more than 1,885 disease-causing mutations of factor VIII gene have been identified. Clinic confers a great challenge for the molecular diagnosis. We aim to make a better strategy for the molecular diagnosis in Hemophilia A. First, factor VIII intron 22 inversion and intron 1 inversion mutations were detected using Inversion-PCR and double-tube multiple PCRs. And then, non-inversion mutations were analyzed by denaturing high performance liquid chromatography and/or direct sequencing. Novel mutations were further analyzed the conservation and 3D structures by a B domain deleted crystallographic model and bioinformatics. Finally, we can indirectly confirm the diagnosis by linkage analysis for the patients with the confusing diagnosis by the techniques mentioned above. Eleven patients with the factor VIII Inv 22 were found, and the remaining 16 patients were found with 11 different mutations, of which 3 was novel mutations affecting A1, B domains and splicing site. Moreover, the prenatal diagnosis was performed on 14 fetuses. Ten fetuses were successfully confirmed to be normal, 1 fetus to be a heterozygote with factor VIII c.3275–3276 ins A and 3 fetuses to be hemizygotes with factor VIII Inv 22 mutation.

Keywords

DHPLC Direct sequencing Genetic diagnosis Hemophilia A Prenatal diagnosis