高等学校化学学报

高等学校化学学报

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聚胸腺嘧啶-铜纳米簇/适体-金纳米粒子荧光探针用于微囊藻毒素-LR传感分析

蒙化1,阿华英1,罗平馨1,张艳丽1,高连训1,王红斌1,杨文荣2,庞鹏飞1   

  1. 1. 云南民族大学 2. 迪肯大学生命与环境科学学院
  • 收稿日期:2025-01-08 修回日期:2025-02-19 出版日期:2025-03-05 发布日期:2025-03-05
  • 通讯作者: 庞鹏飞 E-mail:pfpang@aliyun.com
  • 基金资助:
    国家自然科学基金项目(批准号：21565031, 21665027)，云南省科技厅基础研究计划项目(批准号：202001AT070012)和云南民族大学 研究生科研创新基金项目（批准号：2024SKY135）资助

A Fluorescent Probe for Detection of Microcystin-LR Based on Poly(thymidine)-Copper Nanoclusters and Aptamer-Gold Nanoparticles

MENG Hua1，A Huaying1，LUO Pingxin1，ZHANG Yanli1，GAO Lianxun1，WANG Hongbin1， YANG Wenrong1,2，PANG Pengfei1    

  1. 1. Yunnan Minzu University 2. School of Life and Environmental Sciences, Deakin University
  • Received:2025-01-08 Revised:2025-02-19 Online:2025-03-05 Published:2025-03-05
  • Contact: Pengfei Pang E-mail:pfpang@aliyun.com
  • Supported by:
    Supported by the National Natural Science Foundation of China (Nos. 21665027, 21565031), Basic Research Program Project of Yunnan Provincial Science and Technology Department, China (No. 202001AT070012), and Graduate Research Innovation Fund Project of Yunnan Minzu University, China (No. 2024SKY135)

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摘要： 采用DNA模板法合成聚胸腺嘧啶-铜纳米簇（poly(T)-CuNCs）/适体-金纳米粒子（aptamer-AuNPs）荧光探针，用于微囊藻毒素-LR（MC-LR）高灵敏传感检测. 本工作设计了3条DNA核苷酸链，MC-LR适体链（aptamer）和2条聚胸腺嘧啶单链DNA（poly(T) S1和（poly(T) S2）. 以poly(T) S1和poly(T) S2为模板，利用抗坏血酸（AA）还原Cu2+，合成具有粉红色荧光的poly(T) S1-CuNCs和poly(T) S2-CuNCs. 两端巯基标记的aptamer通过Au-S键与AuNPs相连形成AuNPs-aptamer-AuNPs共轭物，后者与poly(T)-CuNCs杂交形成dsDNA-CuNCs，双链结构中CuNCs与AuNPs之间由于荧光共振能量转移（FRET），导致体系荧光淬灭. 加入目标物MC-LR，由于MC-LR与dsDNA-CuNCs中aptamer特异性结合，引起双链结构解体，poly(T)-CuNCs释放至溶液中，体系荧光恢复. 基于此构建了一种“off-on”型荧光探针用于MC-LR检测，该方法对MC-LR的线性范围为1 ng/L ~ 500 μg/L，检出限为0.3 ng/L (S/N = 3). Poly(T)-CuNCs/aptamer-AuNPs荧光适体探针制备简单、选择性高，可用于实际水样中MC-LR定量分析.

关键词: 微囊藻毒素-LR, 聚胸腺嘧啶-铜纳米簇, 适体-金纳米粒子, 荧光探针

Abstract: A fluorescent probe of poly(thymidine)-copper nanoclusters/aptamer-gold nanoparticles (poly(T)- CuNCs/aptamer-AuNPs) was constructed for highly sensitive sensing detection of microcystin-LR (MC-LR) using DNA template method. Three DNA nucleotides were designed, including MC-LR aptamer, and two poly(thymidine) ssDNA (poly(T) S1 and poly(T) S2). Using poly(T) S1 and poly(T) S2 as templates, poly(T) S1-CuNCs and poly(T) S2-CuNCs with pink fluorescence were synthesized by reduction Cu2+ with ascorbic acid (AA). Aptamer labeled with thiol groups at two both ends, were linked with AuNPs through Au-S bonds to form AuNPs-aptamer-AuNPs bioconjugates. AuNPs-aptamer-AuNPs hybridized with poly(T)-CuNCs to form dsDNA-CuNCs. Fluorescence resonance energy transfer (FRET) occurred between CuNCs and AuNPs in the dsDNA-CuNCs structure, leading to fluorescence quenching of dsDNA-CuNCs. In the presence of target MC-LR, MC-LR specifically bonded with aptamer in dsDNA-CuNCs, resulting in the dissociation of dsDNA structure. The poly(T)-CuNCs were released into the solution, restoring the system fluorescence. An "off-on" type of fluorescent probe was constructed for detection of MC-LR. The linear range for MC-LR detection is 1 ng/L ~ 500 μg/L, with a detection limit of 0.3 ng/L (S/N = 3). The proposed fluorescent aptamer probe possesses the advantages of simple preparation, high selectivity, and can be applied for quantitative detection and analysis of MC-LR in real water samples.

Key words: Microcystin-LR；Poly(thymine)-copper nanoclusters；Aptamer-gold nanoparticles；Fluorescent probe 

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