使用构象敏感凝胶电泳对1型血管性血友病因子基因进行突变分析：荧光技术与手工技术的比较

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.

作者信息

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

机构信息

Academic Unit of Hematology, School of Medicine and Biomedical Science, University of Sheffield, UK.

出版信息

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

DOI:10.3324/haematol.10606

PMID:17488667

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.

摘要

比较了两种构象敏感凝胶电泳技术，即荧光（F-CSGE）和手动（M-CSGE）技术，用于血管性血友病因子基因的突变分析。在7例1型血管性血友病病例以及1例健康对照中，使用56次聚合酶链反应（PCR）扩增该基因的所有52个外显子。每块F-CSGE凝胶分析192个样本，而M-CSGE每块凝胶分析40个样本。125个扩增子在F-CSGE中显示出条带迁移，但在M-CSGE中仅101个显示条带迁移。两种技术均检测到5个突变。F-CSGE检测到45种不同的多态性，而M-CSGE仅检测到39种。F-CSGE具有高通量且比M-CSGE更敏感。