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

Haematologica. 2007 Apr;92(4):550-3. doi: 10.3324/haematol.10606.

Abstract

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.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • DNA / blood
  • DNA / chemistry
  • DNA / genetics*
  • DNA Mutational Analysis / methods*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Exons / genetics
  • Fluorescent Dyes / analysis
  • Fluorometry
  • Heteroduplex Analysis / methods*
  • Humans
  • Nucleic Acid Conformation*
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide
  • Pseudogenes
  • Sensitivity and Specificity
  • von Willebrand Diseases / genetics*
  • von Willebrand Factor / chemistry
  • von Willebrand Factor / genetics*

Substances

  • Fluorescent Dyes
  • von Willebrand Factor
  • DNA