DHPLC screening of cystic fibrosis gene mutations.

摘要: Denaturing high performance liquid chromatography (DHPLC) using ion-pairing reverse phase (IPRPC) columns is a technique for the screening of gene mutations. In order to evaluate potential utility this assay method in clinical laboratory setting, we subjected PCR products 73 CF patients known bear CFTR mutations analytic technique. We used thermal denaturation profile parameters specified by MELT program tool, made available Stanford University. Using strategy, determined an initial sensitivity 90.4% any Most not detected DHPLC under these conditions are alpha-substitutions. This information may eventually help improve algorithm. Increasing column temperatures one or two degrees above those recommended allowed 100% detection tested. By comparing methodology study with recently reported based on Wavemaker 3.4.4 software (Transgenomic, Omaha, NE) [Le Marechal et al., 2001) and previous SSCP analysis [Ravnik-Glavac 1994] emphasized differences similarities refine system discuss relationship alternative approaches. conclude that method, optimized conditions, highly accurate, rapid, efficient detecting find individuals Hum Mutat 19:374-383, 2002. Published 2002 Wiley-Liss, Inc.