High-throughput single strand conformation polymorphism mutation detection by automated capillary array electrophoresis: validation of the method.

摘要: Capillary array electrophoresis (CAE) is a novel technique, which allows for high throughput analysis of DNA fragments. When screening mutations in whole populations or large patient groups it necessary to have robust and well-characterized setups analysis. For large-scale mutation screening, we developed procedures single strand conformation polymorphism (SSCP) assays using CAE (CAE-SSCP) whereby may increase both the sensitivity compared conventional SSCP In this study validated CAE-SSCP by 1) comparing detection slab-gel based with MYH7, MYL2, MYL3 genes encoding sarcomere proteins from patients suffering hypertrophic cardiomyopathy; 2) constructing series 185 mutants having substitution mutations, as well insertion/deletion some combinations these, different sequence contexts four exons positions relative end amplicon (three KCNQ1 gene, cardiac potassium channel, one TNNI3 gene troponin I). The method identified 181 out (98%), data suggest that position fragment had no effect on sensitivity. Analysis specificity showed only very few could not be distinguished each other there were false positives.