© Borgis - Medycyna Rodzinna 4, p. 82-87
*Joanna Skulimowska, Jadwiga Sawecka
Diagnostyka molekularna hemofilii A
Molecular diagnostic of haemophilia A
Zakład Biochemii, Instytut Hematologii i Transfuzjologii w Warszawie
Kierownik Zakładu: dr Urszula Bany-Łaszewicz
Summary
Haemophilia A is a recessive, X-linked bleeding disorder affecting approximately 1/5000 males due to a deficiency or dysfunction of blood coagulation factor VIII. Approximately one third of cases have no prior history of haemophilia A (sporadic haemophilia). The aim of this study is to present a diagnostical strategy for carriership determination and prenatal diagnostics of haemophilia A. The severity of haemophilia A should be determined first because it influences choice of diagnostic strategy. A wide range of different mutation types has been identified, which occur along the entire length of the factor VIII gene. In 20% of haemophilia A cases an inversion in intron 22 of FVIII is the causative mutation, which always leads to severe disease. The second most common mutation in severe disease is the intron 1 inversion mutation, which occurs with a frequency 1-5%. Intron 22 inversion in FVIII gene was directly examined by Southern blot analysis, intron 1 inversion was identified by dual PCR assay. If these mutations are absent, further analysis includes DNA sequencing of those regions in which a mutation is suspected. We used SSCP and HD as screening methods for detecting abnormal fragment. Families with mild and moderate haemophilia A were testing directly by linkage analysis or sequencing. When family has not identified a causative mutation, linkage analysis employing DNA polymorphic markers was used. It is essential to have heterozygous marker in the mother to carry out efficient disease gene tracking. We used dinucleotide polymorphisms in intron 1, 13, 22 and 24.
Key words: factor VIII, haemophilia A, carrier detection, linkage analysis, mutations
Piśmiennictwo
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otrzymano/received: 2008-10-17
zaakceptowano/accepted: 2008-10-24
Adres/address:
*Joanna Skulimowska
Zakład Biochemii,
Instytut Hematologii i Transfuzjologii
ul. Chocimska 5, 00-957 Warszawa
tel.: (0-22) 849 36 51 w 128
e-mail: joskul@ihit.waw.pl
