Detection of heterozygous mutations in BRCA1 using high density oligonucleotide arrays and two-colour fluorescence analysis
Detection of heterozygous mutations in BRCA1 using high density oligonucleotide arrays and two-colour fluorescence analysis
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DOI：
10.1038/ng1296-441
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发表时间：
1996-12-01
期刊：
影响因子：
30.8
通讯作者：
Collins, FS
中科院分区：
文献类型：
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作者：
Hacia, JG;Brody, LC;Collins, FS
在大量患者样本中快速准确地扫描一个大基因的所有可能的杂合突变的能力对医学的未来至关重要。我们设计了由超过96,600个长度为20核苷酸（nt）的寡核苷酸组成的高密度阵列，用于筛选遗传性乳腺癌和卵巢癌基因BRCA1的3.45千碱基（kb）外显子11中的广泛杂合突变。将参考样品和测试样品与这些阵列共杂交，并通过双色分析定量杂交模式的差异。15例已知突变患者样本中有14例被准确诊断，20例对照样本中未发现假阳性突变。8个单核苷酸多态性也很容易检测到。基于DNA芯片的检测可能为高通量、低成本的基因改变检测提供一种有价值的新技术。
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.