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引用本文:   张丽, 潘敏, 邹芷乔, 樊蕾, 刘晓庆. 杂交链式反应介导的发光银纳米簇系统用于miRNA-21的信号放大检测. 分析化学, 2020, 48(9): 1193-1201. doi:  10.19756/j.issn.0253-3820.201144 [复制]

Citation:   ZHANG Li , PAN Min , ZOU Zhi-Qiao , FAN Lei , LIU Xiao-Qing . Hybridization Chain Reaction-Mediated Luminescent Silver Nanocluster System for Amplified Detection of miRNA-21. Chinese Journal of Analytical Chemistry, 2020, 48(9): 1193-1201. doi: 10.19756/j.issn.0253-3820.201144 [复制]

杂交链式反应介导的发光银纳米簇系统用于miRNA-21的信号放大检测

通讯作者:  刘晓庆, xiaoqingliu@whu.edu.cn

收稿日期: 2020-03-19

基金项目: 本文系国家自然科学基金项目(No.81602610)和中央高校基本科研基金项目(No.2042018kf1006)资助

Hybridization Chain Reaction-Mediated Luminescent Silver Nanocluster System for Amplified Detection of miRNA-21

Corresponding author:  LIU Xiao-Qing , xiaoqingliu@whu.edu.cn

Received Date:  2020-03-19

Fund Project:  This work was supported by the National Natural Science Foundation of China (No. 81602610) and the Fundamental Research Funds for the Central Universities, China (No. 2042018kf1006).

MicroRNAs(miRNAs)是多种疾病的生物标志物,同时也能为疾病治疗提供潜在靶点,因此,发展简单、快速、灵敏的miRNAs分析方法具有十分重要的意义。本研究结合杂交链式反应(HCR)高的信号放大能力和以DNA为模板合成的银纳米簇(DNA-AgNCs)优异的发光性能,构建了一种免标记的通用型荧光传感器,实现了miRNA-21的快速灵敏检测。将合成银纳米簇(AgNCs)的DNA模板封闭在HCR的反应物(发夹DNA)中,当存在靶标DNA时,发夹DNA的杂交链式组装反应被引发,释放出大量自由的AgNCs模板序列,进而引发近红外荧光DNA-AgNCs的合成,AgNCs的近红外荧光信号强度与引发链DNA的浓度成正相关。进一步通过在检测系统中引入一个封闭有HCR引发链序列的辅助发卡序列,建立通用型HCR-AgNCs传感分析系统,用于靶核酸分子检测。以miRNA-21为模型分析物,只在miRNA-21存在时,此辅助发卡才能被打开,并生成自由的HCR引发链,进而引发HCR反应和AgNCs的合成。本方法检测miRNA-21的线性范围为250 pmol/L~8 nmol/L,线性方程为(F-F0F0=-0.500+0.235 lg(C(pmol/L))(R2=0.9792),检出限(3σ/S)为19.9 pmol/L。这种基于HCR-AgNCs的检测方法具有免标记、简单、灵敏的优点,可作为通用的核酸传感平台,有望为疾病的临床诊断和治疗提供重要参考。

关键词:   microRNAs, 杂交链式反应, 信号放大, 银纳米簇, DNA组装, 荧光检测
Key words:   MicroRNAs, Hybridization chain reaction, Signal amplification, Silver nanoclusters, DNA assembly, Fluorescence detection
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杂交链式反应介导的发光银纳米簇系统用于miRNA-21的信号放大检测

张丽, 潘敏, 邹芷乔, 樊蕾, 刘晓庆

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