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 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).
Mattocks
C
C
Molecular Genetics Laboratory, Addenbrooke's Hospital, Cambridge, UK. cjm59@cam.ac.uk
Tarpey
P
P
Bobrow
M
M
Whittaker
J
J
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
D018076:DNA, Complementary; D015224:Dideoxynucleosides; D005456:Fluorescent Dyes