Detection of Mutations in Hemophilia A Patients by Chemical Cleavage of Mismatch Method

  • Naushin H. Waseem
  • Richard Bagnall
  • Peter M. Green
  • Francesco Giannelli
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 31)

Abstract

Hemophilia A is an X-linked disorder that leads to a defect in blood coagulation. This is caused by mutations in the factor VIII gene, which results in its activity being reduced or abolished in the blood-clotting cascade. The factor VIII gene is 186 kb long with 26 exons, varying from 69 bp (exon 5) to 3106 bp (exon 14) (1). The factor VIII mRNA is 9028 bases in length with a 7053 nucleotides long coding region (2).

Keywords

Vortex Pyridine Polypropylene MgSO4 Dextran 

References

  1. 1.
    Gitschier, J., Wood, W. I., Goralka, T. M., Wion, K. L., Chen, E. Y., Eaton, D. H., Vehar, G. A., Capon, D. J., and Lawn, R. M. (1984) Characterization of the human factor VIII gene. Nature 312, 326–330.CrossRefPubMedGoogle Scholar
  2. 2.
    Toole, J. J., Knopf, J. L., Wozney, J. M., Sultzman, L. A., Buecker, J. L., Pittman, D. D., Kaufman, R. J., Brown, E., Shoemaker, C., Orr, E. C., Amphlett, G. W., Foster, W. B., Coe, M. L., Knutson, G. J., Fass, D. N., and Hewick, R. M. (1984) Molecular-cloning of a cDNA-encoding human antihemophilic factor. Nature 312, 342–347.CrossRefPubMedGoogle Scholar
  3. 3.
    Higuchi, M., Antonarakis, S. E., Kasch, L., Oldenburg, J., Economou-Petersen, E., Olek, K., Arai, M., Inaba, H., and Kazazian, H. H. (1991) Molecular characterisation of mild-to-moderate hemophelia A: detection of the mutation in 25 of 29 patients by denaturing gel electrophoresis. Proc. Natl. Acad. Sci. USA 88, 8307–8311.CrossRefPubMedGoogle Scholar
  4. 4.
    Orita, M., Iwahana, H., Kanazawa, H., Hayashi, K., and Sekiya, T. (1989) Detection of polymorphisms of human DNA by gel-electrophoresis as single-strand conformation polymorphisms. Proc. Natl. Acad. Sci. USA 86, 2766–2770.CrossRefPubMedGoogle Scholar
  5. 5.
    Naylor, A., Green, P. M., Montandon, A. J., Rizza, C. R., and Giannelli, F. (1991) Detection of three novel mutations in two haemophilia A patients by rapid screening of whole essential region of factor VIII gene. Lancet 337, 635–639.CrossRefPubMedGoogle Scholar
  6. 6.
    Naylor, J. A., Green, P. M., Rizza, C. R., and Giannelli, F. (1992) Factor VIII gene explains all cases of haemophilia A. Lancet 340, 1066,1067.CrossRefPubMedGoogle Scholar
  7. 7.
    Forrest, S. M., Dahl, H. H., Howells, D. N., Dianzani, I., and Cotton, R. G. H. (1991) Mutation detection in phenylketonuria using the chemical cleavage of mismatch method: Importance of using probes from both normal and patient samples. Am. J. Hum. Genet. 49, 175–183.aPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Naushin H. Waseem
    • 1
  • Richard Bagnall
    • 1
  • Peter M. Green
    • 1
  • Francesco Giannelli
    • 1
  1. 1.Paediatric Research Unit, Division of Medical and Molecular Genetics, Prince Philip Research Laboratories, United Medical and Dental Schools of Guy’s and St. Thomas’s Hospitals, Guy’s HospitalUniversity of LondonLondonUK

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