Enhanced detection of mutations in BRCA1 exon 11 using restriction endonuclease fingerprinting-single-strand conformation polymorphism

J Mol Med (Berl). 2000;78(10):580-7. doi: 10.1007/s001090000147.

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.

Publication types

  • Evaluation Study

MeSH terms

  • Breast Neoplasms / genetics*
  • DNA Fingerprinting / methods
  • DNA Restriction Enzymes / metabolism
  • Exons / genetics*
  • Female
  • Genes, BRCA1 / genetics*
  • Genetic Testing / methods*
  • Humans
  • Mutation / genetics*
  • Ovarian Neoplasms / genetics
  • Polymorphism, Single-Stranded Conformational

Substances

  • DNA Restriction Enzymes