高等学校化学学报

高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (5): 1078.doi: 10.7503/cjcu20130029

• 分析化学 • 上一篇    下一篇

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

陈琰1,4, 刘桂锋2, 刘霞3, 张桂珍1, 王振新3   

  1. 1. 吉林大学中日联谊医院中心研究室, 长春 130033;
    2. 吉林大学中日联谊医院放射线科, 长春 130033;
    3. 中国科学院长春应用化学研究所, 电分析国家重点实验室, 长春 130022;
    4. 吉林大学第二医院内分泌科, 长春 130042
  • 收稿日期:2013-01-08 出版日期:2013-05-10 发布日期:2013-05-10
  • 通讯作者: 张桂珍, 女, 教授, 博士生导师, 主要从事糖尿病分子发病机理研究. E-mail: zhangguizhenjl@ yahoo.com.cn E-mail:zhangguizhenjl@yahoo.com.cn
  • 基金资助:

    国家自然科学基金(批准号: 21075118)资助.

Detecting Type 2 Diabetes-related Single Nucleotide Polymorphisms by Microarray

CHEN Yan1,4, LIU Gui-Feng2, LIU Xia3, ZHANG Gui-Zhen1, WANG Zhen-Xin3   

  1. 1. Central Research Department, China-Japan Union Hospital of Jilin University, Changchun 130033, China;
    2. Radiology Department, China-Japan Union Hospital of Jilin University, Changchun 130033, China;
    3. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun 130022, China;
    4. Endocrinology Department, The Second Hospital of Jilin University, Changchun 130042, China
  • Received:2013-01-08 Online:2013-05-10 Published:2013-05-10
  • Contact: CHEN Yan E-mail:zhangguizhenjl@yahoo.com.cn

PDF (PC)

被引次数

4

摘要：

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.

关键词: 寡核苷酸微阵列, 2型糖尿病, 等位基因频率, 核苷酸探针

Abstract:

A three-dimensional(3D) oligonucleotide microarray-based fluorescent assay was developed for screening specific oligonucleotide probes for type 2 diabetes detection. The 3D oligonucleotide microarray was fabricated on the home-made fourth-generation(G4) NH2-terminated poly(amidoamine) dendrimers(PAMAM) 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) oligonucleotides, 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.

Key words: Oligonucleotide microarray, Type 2 diabetes, Allele frequency, Oligonucleotide probe

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