Comparative sequence analysis (CSA): A new sequence-based method for the identification and characterization of mutations in DNA

Direct sequencing analysis is largely used to confirm and characterize muta tions previously detected by more rapid tests. We have developed a method-C omparative Sequence Analysis (CSA) - that simplifies the analysis of sequen cing data facilitating its use as a first screen for mutation detection. Se quence data were split into their component electrophoretograms and the use of a size standard enabled equivalent traces from different individuals to be overlaid. This allowed simple and rapid visual analysis of the results, Using this technique in a blind study, we tested 576 samples for mutations in the Von Hippel-Lindau tumor suppresser gene, VHL. We were able to ident ify and characterize all 78 known mutations present within the sample set ( 100% sensitivity and specificity). Hum Mutat 16:437-443, 2000. (C) 2000 Wil ey-Liss, Inc.