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.

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.

• Authors: Mohammad Hashemi Soteh, Ian R Peake, Luke Marsden, John Anson, Javier Batlle, Dominique Meyer, Edith Fressinaud, Claudine Mazurier, Jenny Goudemand, Jeroen Eikenboom, Anne Goodeve, MCMDM-1VWD Study Group
• Journal: Haematologica (Haematologica) Vol. 92 Issue 4 Pg. 550-3 (Apr 2007) ISSN: 1592-8721 [Electronic] Italy
• PMID: 17488667 (Publication Type: Comparative Study, Evaluation Study, Journal Article)

Chemical References

• Fluorescent Dyes
• von Willebrand Factor
• DNA

Topics

• DNA (blood, chemistry, 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, genetics)