银染增强的纳米金标记探针对微量核酸的检测.pdfVIP

ISSN 中国生物化学与分子生物学报 2006 年 11 月 CN 　113870Q Chinese Journal of Biochemistry and Molecular Biology 22 ( 11) :919～923 ·技术与方法· 银染增强的纳米金探针技术检测微量核酸 赵立凡 , 　李柏生 , 　黑笑涵 , 　李晓波 , 　李媛媛 , 　徐顺清 (华中科技大学同济医学院 公共卫生学院 环境医学研究所 环境与健康教育部重点实验室 ,武汉　430030) 摘要 　利用银染增强的纳米金技术建立了一种简单快速的核酸定量方法. 该方法基于纳米金与烷 巯基修饰的寡核苷酸分子共价键合作用 ,将纳米微粒报告基团标记在与靶核酸一端序列互补的寡 核苷酸上 , 同时生物素化修饰另一端互补序列. 靶核酸与两段寡核苷酸探针杂交后 ,借亲和素固定 在酶标板孔内 ,通过纳米金催化的银染放大效应产生高灵敏的识别信号 ,适时记录其吸光度值从而 实现核酸分子的定量. 该检测方法检测单链核酸分子的灵敏度达 01 fmolL ,双链分子为 10 fmolL. 关键词 　纳米金 ; 核酸检测 ; 银染增强 中图分类号 　R115 A Colorimetric Polynucleotides Detection Method Based on Nanoparticle Probes with Silver Staining Enhancement ZHAO LiFan , LI BoSheng , HEI XiaoHan , LI XiaoBo , LI YuanYuan ,XU ShunQing ( MOE Key Laboratory of Environment and Health , School of Public Health , Tongji Medical College of Huazhong University of Science and Technology , Wuhan 　430030 , China) Abstract 　A simple gold nanoparticlebased method to detect micro amount of polynucleotide molecules with silver staining enhancement is established. Based on the covalence action of thiolmodified oligonucleotides with nanoparticles , the nanoparticles was utilized as biomarkers to detect trace amount of polynucleotides. Two kinds of probes , which were respectively complementary to one half of target polynucleotides , were labeled respectively with biotins and nanoparticles. The hybridization products of target polynucleotides with these two kinds of probes were immobilized to microplates via avidinbiotin system , and the signals of gold nanopart