Nature Genetics
14, 441 - 447 (1996)
doi:10.1038/ng1296-441
Detection of heterozygous mutations in BRCA1 using high density oligonucleotide arrays and two−colour fluorescence analysisJoseph G. Hacia1, Lawrence C. Brody1, Mark S. Chee2, Stephen P.A. Fodor2
& Francis S. Collins1, 3
1National Center for Human Genome Research, Building 49/3A14, National Institutes of Health, Bethesda, Maryland 20892, USA
2Affymetrix, 3380 Central Expressway, Santa Clara, California 95051, USA
3Correspondence should be addressed to F.S.C. 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.
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