Lovmar Lovisa, Syvänen Ann-Christine
Molecular Medicine, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Hum Mutat. 2006 Jul;27(7):603-14. doi: 10.1002/humu.20341.
In many situations there may not be sufficient DNA collected from patient or population cohorts to meet the requirements of genome-wide analysis of SNPs, genomic copy number polymorphisms, or acquired copy number alternations. When the amount of available DNA for genotype analysis is limited, high performance whole-genome amplification (WGA) represents a new development in genetic analysis. It is especially useful for analysis of DNA extracted from stored histology slides, tissue samples, buccal swabs, or blood stains collected on filter paper. The multiple displacement amplification (MDA) method, which relies on isothermal amplification using the DNA polymerase of the bacteriophage phi29, is a recently developed technique for high performance WGA. This review addresses new trends in the technical performance of MDA and its applications to genetic analyses. The main challenge of WGA methods is to obtain balanced and faithful replication of all chromosomal regions without the loss of or preferential amplification of any genomic loci or allele. In multiple comparisons to other WGA methods, MDA appears to be most reliable for genotyping, with the most favorable call rates, best genomic coverage, and lowest amplification bias.
在许多情况下，从患者或人群队列中收集的DNA可能不足以满足对单核苷酸多态性（SNP）、基因组拷贝数多态性或获得性拷贝数改变进行全基因组分析的要求。当用于基因型分析的可用DNA量有限时，高性能全基因组扩增（WGA）代表了遗传分析中的一项新进展。它对于分析从储存的组织学切片、组织样本、口腔拭子或滤纸上收集的血渍中提取的DNA特别有用。多重置换扩增（MDA）方法依赖于使用噬菌体phi29的DNA聚合酶进行等温扩增，是最近开发的一种用于高性能WGA的技术。本文综述了MDA技术性能的新趋势及其在遗传分析中的应用。WGA方法的主要挑战是获得所有染色体区域的平衡且忠实的复制，而不会丢失或优先扩增任何基因组位点或等位基因。在与其他WGA方法的多次比较中，MDA在基因分型方面似乎最可靠，具有最有利的检出率、最佳的基因组覆盖范围和最低的扩增偏差。
