Comparative genomic hybridization, loss of heterozygosity, and DNA sequence analysis of single cells
Comparative genomic hybridization, loss of heterozygosity, and DNA sequence analysis of single cells
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DOI：
10.1073/pnas.96.8.4494
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发表时间：
1999-04-13
影响因子：
11.1
通讯作者：
Riethmüller, G
中科院分区：
文献类型：
--
作者：
Klein, CA;Schmidt-Kittler, O;Riethmüller, G
描述了用于从单个真核细胞中全局扩增DNA的PCR策略，其使得能够对整个基因组进行全面分析。通过比较基因组杂交，不仅可以可靠地检索总DNA拷贝数变异，例如分别在单个雄性细胞和唐氏综合征患者细胞中的单体X和三体21，而且可以可靠地检索人类肿瘤细胞的多个缺失和扩增特征。作为暴露于选择压力的异质细胞群体的模型，我们研究了从癌症患者骨髓中分离的单个微转移细胞。所观察到的一致模式的比较基因组杂交数据，杂合性丢失，和突变，通过测序检测证明了该技术的保真度，并证明其用于评估复杂人群中的遗传变异的克隆进化的有用性。
A PCR strategy is described for global amplification of DNA from a single eukaryotic cell that enables the comprehensive analysis of the whole genome. By comparative genomic hybridization, not only gross DNA copy number variations, such as monosomic X and trisomic 21 in single male cells and cells from Down's syndrome patients, respectively, but multiple deletions and amplifications characteristic for human tumor cells are reliably retrieved. As a model of heterogeneous cell populations exposed to selective pressure, we have studied single micrometastatic cells isolated from bone marrow of cancer patients. The observed congruent pattern of comparative genomic hybridization data, loss of heterozygosity, and mutations as detected by sequencing attests to the technique's fidelity and demonstrates its usefulness for assessing clonal evolution of genetic variants in complex populations.