基于纳米金比色检测NOS1AP基因单碱基突变

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (10): 1965.

基于纳米金比色检测NOS1AP基因单碱基突变

郭青川1,2, 王祥1, 娄新徽1, 毛红菊1, 贾杰3, 金庆辉1, 赵建龙1, 张峰3

1. 中国科学院上海微系统与信息技术研究所, 上海 200050;
2. 新乡医学院临床三系康复医学科, 新乡 453003;
3. 复旦大学附属华山医院康复医学科, 上海 200040

收稿日期:2009-12-30 出版日期:2010-10-10 发布日期:2010-10-10
通讯作者: 毛红菊, E-mail: hjmao@mail.sim.ac.cn; 贾杰, E-mail: drjiajie@163.com
基金资助:
国家重大科学计划项目(批准号: 2009ZX10004-105)和上海市科委纳米专项基金(批准号: 0952nm05700)资助.

Label-free Colorimetric Assay Based on Gold Nanoparticles in Detection of NOS1AP with Single Nucleotide Polymorphism

GUO Qing-Chuan1,2, WANG Xiang1, LOU Xin-Hui1, MAO Hong-Ju1*, JIA Jie3*, JIN Qing-Hui1, ZHAO Jian-Long1, ZHANG Feng3

1. Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China;
2. Department of Rehabilitation, Third Hospital, Xinxiang Medical College, Xinxiang 453003, China;
3. Department of Rehabilitation, Huashan Hospital, Fudan University, Shanghai 200040, China

Received:2009-12-30 Online:2010-10-10 Published:2010-10-10
Contact: MAO Hong-Ju, E-mail: hjmao@mail.sim.ac.cn; JIA Jie, E-mail: drjiajie@163.com
Supported by:
国家重大科学计划项目(批准号: 2009ZX10004-105)和上海市科委纳米专项基金(批准号: 0952nm05700)资助.

摘要/Abstract

摘要： 基于单、双链DNA与纳米金颗粒间的不同静电作用, 建立了一种基于颜色反应检测NOS1AP基因单碱基突变的方法. 根据NOS1AP基因的单碱基多态位点设计检测探针、互补靶序列及带有单碱基突变序列寡核苷酸DNA. 室温下, 检测探针分别与互补序列、单碱基突变序列在缓冲液中进行杂交, 再分别加入纳米金溶液以及NaCl溶液. 用肉眼可以观察到纳米金溶液在两种不同杂交溶液中产生明显不同的颜色变化. 这种变化可通过紫外-可见分光光度计测定纳米金溶液的紫外吸收峰值的变化来证实. 实验结果表明, 纳米金溶液在一定浓度NaCl存在的条件下, 对互补双链NOS1AP DNA及单碱基突变NOS1AP DNA呈现出不同的颜色反应及紫外吸收光谱的改变. 此方法可望用于相关疾病的医学诊断及单碱基突变的检测.

关键词: 纳米金, NOS1AP, 单核苷酸多态性, 比色检测

Abstract: A gold nanoparticles’ colorimetry-based assay for detection of NOS1AP gene with single nucleotide polymorphism was established, which profited from the dissimilar electrostatic interactions between single-stranded oligonucleotides and double-stranded oligonucleotides. The detection probes, the oligonucleotides with their perfectly complementary sequence, and the sequence with a single mismatched base were designed according to the NOS1AP gene sequence. The detection probes were hybridized with their complementary oligonucleotides and single-base-mismatch oligonucleotides, respectively. Then, gold nanoparticles solutions and NaCl solutions were added, respectively. Different color appeared obviously by naked eye which showed an evident change in UV-Vis absorption spectroscopy similarly. The results indicate that with the presence of the appropriate concentration of NaCl, the gold nanoparticles solutions with double-stranded oligonucleotides are distinguished from gold nanoparticles solutions with single mismatched oligonucleotides by colormetric assay. This method enables a simple, rapid and efficient detection that could have potential and possible application prospects in medical diagnosis.

Key words: Gold nanoparticle, NOS1AP, Single nucleotide polymorphism, Colorimetric assay

TrendMD:

郭青川, 王祥, 娄新徽, 毛红菊, 贾杰, 金庆辉, 赵建龙, 张峰. 基于纳米金比色检测NOS1AP基因单碱基突变. 高等学校化学学报, 2010, 31(10): 1965.

GUO Qing-Chuan, WANG Xiang, LOU Xin-Hui, MAO Hong-Ju*, JIA Jie*, JIN Qing-Hui, ZHAO Jian-Long, ZHANG Feng. Label-free Colorimetric Assay Based on Gold Nanoparticles in Detection of NOS1AP with Single Nucleotide Polymorphism. Chem. J. Chinese Universities, 2010, 31(10): 1965.

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