title: Constant denaturant gel electrophoresis (CDGE) in BRCA1 mutation screening.
abstract: Screening for mutations in the breast and ovarian cancer susceptibility gene, BRCA1, is complicated by the wide spectrum of mutations found in this large gene. In the present study a constant denaturant gel electrophoresis (CDGE) mutation screening strategy was established for approximately 80% of the genomic coding sequence (exons 2, 11, 13-16, 20, 24). This strategy was applied to screen genomic DNA from 50 familial breast and/or ovarian cancer patients who had previously been examined for BRCA1 mutations by SSCP. A total of 14 carriers of 12 distinct disease-associated mutations and 7 carriers of 6 distinct rare substitutions leading to amino acid substitutions were identified. The SSCP failed to detect 40% of the different deletions/insertions (4/10) and 75% (6/8) of the different base substitutions leading to terminating codons or rare amino acid changes. SSCP did, however, identify one rare base substitution that could not be detected in the CDGE screening. To evaluate the CDGE mutation screening strategy further, 25 unrelated patients from Norwegian breast and/or ovarian cancer families were examined for BRCA1 mutations using a combined genomic DNA/cDNA approach covering the entire coding sequence of the gene. A total of six mutation carriers were detected, all of whom had cases of ovarian cancer in their families. Three patients from independent families carried an 1135insA mutation in exon 11, two others had a Gly484ter and an 1675delA mutation, respectively, and the sixth carried a splice mutation (5194-2 a-->c) causing deletion of exon 18. CDGE may become an efficient tool in diagnostic and population based screening for BRCA1 mutations.
authors.affiliation: Department of Genetics, Institute for Cancer Research, Norwegian Radium Hospital, Oslo.
authors.forename: T I
authors.initials: TI
authors.lastname: Andersen
authors.forename: H G
authors.initials: HG
authors.lastname: Eiken
authors.forename: F
authors.initials: F
authors.lastname: Couch
authors.forename: G
authors.initials: G
authors.lastname: Kaada
authors.forename: M
authors.initials: M
authors.lastname: Skrede
authors.forename: H
authors.initials: H
authors.lastname: Johnsen
authors.forename: T A
authors.initials: TA
authors.lastname: Aloysius
authors.forename: K M
authors.initials: KM
authors.lastname: Tveit
authors.forename: L
authors.initials: L
authors.lastname: Tranebjaerg
authors.forename: A
authors.initials: A
authors.lastname: Dørum
authors.forename: P
authors.initials: P
authors.lastname: Møller
authors.forename: B L
authors.initials: BL
authors.lastname: Weber
authors.forename: A L
authors.initials: AL
authors.lastname: Børresen-Dale
mesh_terms: D019313:BRCA1 Protein / Q000235:genetics*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics; D004252:DNA Mutational Analysis; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005260:Female; D006580:Genetic Carrier Screening; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D009664:Norway; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics; D011110:Polymorphism, Genetic / Q000235:genetics; D013696:Temperature
chemical_list: D019313:BRCA1 Protein; D017931:DNA Primers