应用微阵列芯片检测与2型糖尿病相关的单核苷酸多态性

Detecting Type 2 Diabetes-related Single Nucleotide Polymorphisms by Microarray

基于三维(3D)寡核苷酸微阵列芯片的荧光检测法,研制了一种用于筛选能检测2型糖尿病的特定寡核苷酸探针.使用第4代(G4)聚(酰胺-胺)(PAMAM)树枝状大分子修饰的载玻片为基底,以氨基修饰的寡核苷酸为固定探针构建3D寡核苷酸微阵列芯片.采用荧光化合物Cy5修饰的寡核苷酸为检测探针获得荧光信号.以2型糖尿病易感基因TCF7L2的rs7903146位点为研究对象,通过对含有16种(8对)寡核苷酸的寡核苷酸文库的筛选,获得了1对能用于2型糖尿病检测的寡核苷酸探针.通过单核苷酸多态性和等位基因分析证明,该寡核苷酸探针对靶标寡核苷酸检测具有高特异性,并能准确检测低至2%的等位基因频率.

A three-dimensional （3D） oligonucleotide microarray-based fluorescent assay was developed for screening specific oligonucleotide probes for type 2 diabetes detection. The 3D oligonucleotide mieroarray was fabricated on the home-made fourth-generation （ G4 ） NH2-terminated poly （amidoamine） dendrimers （ PAM- AM） modified substrate. Assay for rs7903146 site of the type 2 diabetes related gene TCF7L2 was developed. The utility of this assay was demonstrated with an oligonucleotide library, a collection of 16 （8 pairs） oligonu- cleotides, and one specific oligonucleotide probe pair for type 2 diabetes was obtained. Taking advantage of the high specificity of the probe pair, the allele frequency as low as 2% can be accurately determined.