Pomeroy Robert, Duncan George, Sunar-Reeder Bulbin, Ortenberg Elen, Ketchum Melba, Wasiluk Hannah, Reeder Dennis
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, 92093, USA.
Anal Biochem. 2009 Dec 1;395(1):61-7. doi: 10.1016/j.ab.2009.07.041. Epub 2009 Jul 30.
Single nucleotide polymorphism (SNP) analysis of human DNA for the purpose of identification has some promising attributes. The question of approach is critical to the eventual adoption of this technology. The use of a low-volume open array platform was tested with a small selected set of eight SNP primers that have a low F(ST) (the proportion of the total genetic variance contained in a subpopulation [S subscript] relative to the total genetic variance [T subscript]) in human populations. Because multiple SNPs must be interrogated, issues concerning DNA concentration, total DNA, and whole genome amplification were investigated. Excellent correlations were obtained for seven of the eight SNP assays on a set of DNA samples of known configuration over a broad concentration range spanning 25-150ng/microl in blind studies. These seven SNP assays were then applied to 39 DNA samples in a population from southern India. These SNPs were sufficient to individualize each member of this sample population. In a paternity study, these same SNPs showed clear parental relationships. For low amounts of genomic DNA, the use of a commercially available whole genome amplification kit showed promise for genotyping sub-nanogram samples. Discrimination against nonhuman DNA was also demonstrated successfully. Because of the very low quantities of reagents used in the assay, the cost per test becomes reasonably inexpensive. Overall, using commercially available SNP assays, the OpenArray platform showed excellent promise as a highly automated, low-volume, high-throughput system for SNP analysis with potential applications to relevant forensic analyses such as identification and paternity.
用于身份识别目的的人类DNA单核苷酸多态性（SNP）分析具有一些有前景的特性。方法问题对于该技术最终能否被采用至关重要。使用一个低通量开放阵列平台，对一小部分经过挑选的、在人类群体中具有低F(ST)（亚群体[S下标]中所含总遗传变异相对于总遗传变异[T下标]的比例）的八个SNP引物进行了测试。由于必须对多个SNP进行检测，因此研究了与DNA浓度、总DNA和全基因组扩增有关的问题。在盲法研究中，对于一组已知配置的DNA样本，在25 - 150ng/微升的广泛浓度范围内，八个SNP检测中的七个获得了极好的相关性。然后将这七个SNP检测应用于来自印度南部一个群体的39个DNA样本。这些SNP足以区分该样本群体中的每个成员。在一项亲子鉴定研究中，同样的这些SNP显示出明确的亲子关系。对于少量的基因组DNA，使用市售的全基因组扩增试剂盒显示出对亚纳克级样本进行基因分型的前景。对非人类DNA的区分也得到了成功证明。由于检测中使用的试剂数量非常少，每次检测的成本变得相当低廉。总体而言，使用市售的SNP检测，OpenArray平台作为一个高度自动化、低通量、高通量的SNP分析系统显示出极好的前景，具有应用于身份识别和亲子鉴定等相关法医分析的潜力。
