Detection of heterozygous mutations in BRCA1 using high density oligonucleotide arrays and two-colour fluorescence analysis

Nat Genet. 1996 Dec;14(4):441-7. doi: 10.1038/ng1296-441.

Abstract

The ability to scan a large gene rapidly and accurately for all possible heterozygous mutations in large numbers of patient samples will be critical for the future of medicine. We have designed high-density arrays consisting of over 96,600 oligonucleotides 20-nucleotides (nt) in length to screen for a wide range of heterozygous mutations in the 3.45-kilobases (kb) exon 11 of the hereditary breast and ovarian cancer gene BRCA1. Reference and test samples were co-hybridized to these arrays and differences in hybridization patterns quantitated by two-colour analysis. Fourteen of fifteen patient samples with known mutations were accurately diagnosed, and no false positive mutations were identified in 20 control samples. Eight single nucleotide polymorphisms were also readily detected. DNA chip-based assays may provide a valuable new technology for high-throughput cost-efficient detection of genetic alterations.

MeSH terms

  • BRCA1 Protein / genetics*
  • DNA Probes*
  • False Negative Reactions
  • Feasibility Studies
  • Fluorescein
  • Fluoresceins
  • Fluorescence
  • Genetic Carrier Screening
  • Heterozygote*
  • Humans
  • Mutation*
  • Nucleic Acid Hybridization
  • Sensitivity and Specificity

Substances

  • BRCA1 Protein
  • DNA Probes
  • Fluoresceins
  • Fluorescein