title: Comparative sequence analysis (CSA): a new sequence-based method for the identification and characterization of mutations in DNA.
abstract: Direct sequencing analysis is largely used to confirm and characterize mutations previously detected by more rapid tests. We have developed a method-Comparative Sequence Analysis (CSA)-that simplifies the analysis of sequencing data facilitating its use as a first screen for mutation detection. Sequence 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 suppressor gene, VHL. We were able to identify and characterize all 78 known mutations present within the sample set (100% sensitivity and specificity).
authors.affiliation: Molecular Genetics Laboratory, Addenbrooke's Hospital, Cambridge, UK. cjm59@cam.ac.uk
authors.forename: C
authors.initials: C
authors.lastname: Mattocks
authors.forename: P
authors.initials: P
authors.lastname: Tarpey
authors.forename: M
authors.initials: M
authors.lastname: Bobrow
authors.forename: J
authors.initials: J
authors.lastname: Whittaker
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D015224:Dideoxynucleosides / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism; D016368:Frameshift Mutation; D016147:Genes, Tumor Suppressor / Q000235:genetics; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods*; D016037:Single-Blind Method; D012984:Software; D006623:von Hippel-Lindau Disease / Q000175:diagnosis / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D015224:Dideoxynucleosides; D005456:Fluorescent Dyes