Bansal Aruna, van den Boom Dirk, Kammerer Stefan, Honisch Christiane, Adam Gail, Cantor Charles R, Kleyn Patrick, Braun Andi
SEQUENOM-Gemini Ltd., 162 Cambridge Science Park, Milton Road, Cambridge, CB4 0GH England.
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16871-4. doi: 10.1073/pnas.262671399. Epub 2002 Dec 10.
With an ever-increasing resource of validated single-nucleotide polymorphisms (SNPs), the limiting factors in genome-wide association analysis have become genotyping capacity and the availability of DNA. We provide a proof of concept of the use of pooled DNA as a means of efficiently screening SNPs and prioritizing them for further study. This approach reduces the final number of SNPs that undergo full, sample-by-sample genotyping as well as the quantity of DNA used overall. We have examined 15 SNPs in the cholesteryl ester transfer protein (CETP) gene, a gene previously demonstrated to be associated with serum high-density lipoprotein cholesterol levels. The SNPs were amplified in two pools of DNA derived from groups of individuals with extremely high and extremely low serum high-density lipoprotein cholesterol levels, respectively. P values <0.05 were obtained for 14 SNPs, supporting the described association. Genotyping of the individual samples showed that the average margin of error in frequency estimate was approximately 4% when pools were used. These findings clearly demonstrate the potential of pooling techniques and their associated technologies as an initial screen in the search for genetic associations.
随着经过验证的单核苷酸多态性（SNP）资源不断增加，全基因组关联分析中的限制因素已变为基因分型能力和DNA的可获得性。我们提供了一个概念验证，即使用混合DNA作为有效筛选SNP并将其优先用于进一步研究的一种手段。这种方法减少了进行逐个样本全面基因分型的SNP最终数量以及总体使用的DNA量。我们检测了胆固醇酯转运蛋白（CETP）基因中的15个SNP，该基因先前已被证明与血清高密度脂蛋白胆固醇水平相关。这些SNP分别在来自血清高密度脂蛋白胆固醇水平极高和极低个体组的两池DNA中进行扩增。14个SNP的P值<0.05，支持了所描述的关联。对个体样本进行基因分型表明，使用混合样本时频率估计的平均误差幅度约为4%。这些发现清楚地证明了混合技术及其相关技术作为寻找基因关联的初始筛选方法的潜力。
