高等学校化学学报 ›› 2015, Vol. 36 ›› Issue (6): 1061-1066.doi: 10.7503/cjcu20150016

• 分析化学 • 上一篇    下一篇

基于聚多巴胺纳米粒子的荧光共振能量转移法检测microRNA

马立娜1, 柳扶摇1,2, 高嘉雪1,2, 王振新1()   

  1. 1. 中国科学院长春应用化学研究所, 电分析化学国家重点实验室, 长春130022
    2. 中国科学院大学, 北京 100049
  • 收稿日期:2015-01-07 出版日期:2015-06-10 发布日期:2015-05-05
  • 作者简介:联系人简介: 王振新, 男, 博士, 研究员, 博士生导师, 主要从事高通量分析和生物纳米技术方面的研究. E-mail:wangzx@ciac.ac.cn
  • 基金资助:
    国家自然科学基金(批准号: 21127010)资助

Polydopamine Nanoparticle-based Fluorescence Resonance Energy Transfer Assay for microRNA Detection

MA Lina1, LIU Fuyao1,2, GAO Jiaxue1,2, WANG Zhenxin1,*()   

  1. 1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry,Chinese Academy of Sciences, Changchun 130022, China
    2. University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2015-01-07 Online:2015-06-10 Published:2015-05-05
  • Contact: WANG Zhenxin E-mail:wangzx@ciac.ac.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(No.21127010)

摘要:

基于聚多巴胺纳米粒子(PDA NPs)对Cy5标记单链DNA(Cy5-ssDNA)探针的荧光猝灭效应以及脱氧核糖核酸酶Ⅰ(DNaseⅠ)选择性切割DNA/RNA杂合结构中单链DNA的特性, 建立了一种用于微小核糖核酸(miRNA)检测的新型恒温信号放大方法. 在优化的实验条件下, 体系的相对荧光强度(FR)与miR-21浓度的对数值成正比; 对miR-21检测的线性范围为10 pmol/L~100 nmol/L, 检出限达7 pmol/L. 血清加标实验结果表明, 该方法可用于生理环境下miR-21的检测.

关键词: 聚多巴胺纳米粒子, 微小核糖核酸, 脱氧核糖核酸酶Ⅰ, 荧光共振能量转移

Abstract:

A simple, rapid and low-cost polydopamine nanoparticle(PDA NP)-based fluorescence resonance energy transfer(FRET) assay with deoxyribonucleaseⅠ(DNaseⅠ)-assisted target recycling signal amplification was developed for microRNA(miRNA) detection. Under the optimized experimental conditions, the recovery ratio of Cy5 fluorescence intensity increased linearly with logarithm of miR-21 concentration from 10 pmol/L to 100 nmol/L, with a detection limit of 7 pmol/L. Moreover, satisfactory results were obtained while the proposed method was applied to detect miRNA in 10% human serum.

Key words: Polydopamine nanoparticle, microRNA, DNaseⅠ, Fluorescence resonance energy transfer

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