title: Enhanced detection of mutations in BRCA1 exon 11 using restriction endonuclease fingerprinting-single-strand conformation polymorphism.
abstract: A novel approach to mutation screening in the large exon 11 (comprising 3427 bp) of the human BRCA1 gene is presented. Restriction endonuclease fingerprinting single-strand conformation polymorphism (REF-SSCP) is based on repeated detection of DNA sequence variants in different restriction endonuclease fragments, and we evaluated the method using blood samples from 25 Norwegian patients with hereditary breast/ovarian cancer. We compared REF-SSCP to constant denaturant gel electrophoresis (CDGE) and to the protein truncation test (PTT). REF-SSCP detected 12 different DNA variants. Four of these were not detected by CDGE, and only one variant detected by CDGE was missed by REF-SSCP. PTT detected 4 of these 13 variants. REF-SSCP was subsequently applied to a second patient series (Swedish, n=20). A total of 14 different DNA variants were detected by REF-SSCP, 6 of which were truncating mutations (PTT detected only 4). Nonsense and frameshift mutations that are putative breast/ovarian cancer mutations, were detected in 7 of the 25 Norwegian and 9 of the 20 Swedish patients.
authors.affiliation: Center for Molecular Medicine, Haukeland University Hospital, Bergen, Norway.
authors.forename: A
authors.initials: A
authors.lastname: Jugessur
authors.forename: P
authors.initials: P
authors.lastname: Frost
authors.forename: T I
authors.initials: TI
authors.lastname: Andersen
authors.forename: S
authors.initials: S
authors.lastname: Steine
authors.forename: A
authors.initials: A
authors.lastname: Lindblom
authors.forename: A L
authors.initials: AL
authors.lastname: Børresen-Dale
authors.forename: H G
authors.initials: HG
authors.lastname: Eiken
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D005260:Female; D019398:Genes, BRCA1 / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D009154:Mutation / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000235:genetics; D018807:Polymorphism, Single-Stranded Conformational
chemical_list: D004262:DNA Restriction Enzymes