Haematologica (Apr 2007)

Mutational analysis of the von Willebrand factor gene in type 1 von Willebrand disease using conformation sensitive gel electrophoresis: a comparison of fluorescent and manual techniques

  • Mohammad Hashemi Soteh,
  • Ian R Peake,
  • Luke Marsden,
  • John Anson,
  • Javier Batlle,
  • Dominique Meyer,
  • Edith Fressinaud,
  • Claudine Mazurier,
  • Jenny Goudemand,
  • Jeroen Eikenboom,
  • Anne Goodeve

DOI
https://doi.org/10.3324/haematol.10606
Journal volume & issue
Vol. 92, no. 4

Abstract

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Two versions of conformation sensitive gel electrophoresis, fluorescent (F-CSGE) and manual (M-CSGE) techniques, were compared for mutation analysis of the von Willebrand factor gene. 56 PCRs were used to amplify all 52 exons of the gene in seven type 1 von Willebrand disease cases, plus a healthy control. One hundred and ninety-two samples were analyzed on each F-CSGE gel, compared with 40 on M-CSGE. 125 amplicons revealed bandshifts using F-CSGE, but only 101 by M-CSGE. Five mutations were detected by both techniques. F-CSGE detected 45 different polymorphisms whereas M-CSGE detected only 39. F-CSGE is high-throughput and more sensitive than M-CSGE.