School of Public Health, University of California, Berkeley, California 94720, USA.
Environ Mol Mutagen. 2010 Oct-Dec;51(8-9):851-70. doi: 10.1002/em.20630.
Technical advances have improved the capacity to detect and quantify genetic variants, providing novel methods for the detection of rare mutations and for better understanding the underlying environmental factors and biological mechanisms contributing to mutagenesis. The polymerase chain reaction (PCR) has revolutionized genetic testing and remains central to many of these new techniques for mutation detection. Millions of genetic variations have been discovered across the genome. These variations include germline mutations and polymorphisms, which are inherited in a Mendelian manner and present in all cells, as well as acquired, somatic mutations that differ widely by type and size [from single-base mutations to whole chromosome rearrangements, and including submicroscopic copy number variations (CNVs)]. This review focuses on current methods for assessing acquired somatic mutations in the genome, and it examines their application in molecular epidemiology and sensitive detection and analysis of disease. Although older technologies have been exploited for detecting acquired mutations in cancer and other disease, the high-throughput and high-sensitivity offered by next-generation sequencing (NGS) systems are transforming the discovery of disease-associated acquired mutations by enabling comparative whole-genome sequencing of diseased and healthy tissues from the same individual. Emerging microfluidic technologies are beginning to facilitate single-cell genetic analysis of target variable regions for investigating cell heterogeneity within tumors as well as preclinical detection of disease. The technologies discussed in this review will significantly expand our knowledge of acquired genetic mutations and causative mechanisms.
技术的进步提高了检测和量化基因突变的能力，为检测罕见突变提供了新的方法，并有助于更好地了解导致突变的潜在环境因素和生物学机制。聚合酶链反应 (PCR) 彻底改变了基因检测，仍然是许多这些新的突变检测技术的核心。在整个基因组中发现了数百万种遗传变异。这些变异包括种系突变和多态性，它们以孟德尔方式遗传，存在于所有细胞中，以及获得性、体细胞突变，其类型和大小差异很大[从单个碱基突变到整个染色体重排，包括亚微观拷贝数变异 (CNV)]。这篇综述重点介绍了目前评估基因组中获得性体细胞突变的方法，并探讨了它们在分子流行病学中的应用以及疾病的敏感检测和分析。尽管旧技术已被用于检测癌症和其他疾病中的获得性突变，但下一代测序 (NGS) 系统提供的高通量和高灵敏度正在通过能够对来自同一个体的患病和健康组织进行全基因组比较测序来改变疾病相关获得性突变的发现。新兴的微流控技术开始促进针对靶变区的单细胞遗传分析，以研究肿瘤内的细胞异质性以及临床前疾病的检测。本综述中讨论的技术将极大地扩展我们对获得性遗传突变和因果机制的认识。
