基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原

引用本文: 李紫滢, 李德燕, 杨建梅, 胡蓉, 杨通, 杨云慧. 基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原[J]. 分析化学, 2022, 50(7): 1032-1040. doi: 10.19756/j.issn.0253-3820.210686 shu

Citation: LI Zi-Ying, LI De-Yan, YANG Jian-Mei, HU Rong, YANG Tong, YANG Yun-Hui. Fluorescence Resonance Energy Transfer-DNA Nanomachine-based Cycling Signal Amplified Strategy for Detection of Prostate Specific Antigen[J]. Chinese Journal of Analytical Chemistry, 2022, 50(7): 1032-1040. doi: 10.19756/j.issn.0253-3820.210686 shu

基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原

云南师范大学化学化工学院, 昆明 650500

通讯作者: 杨通,E-mail:yangtong2018@ynnu.edu.cn; 杨云慧,E-mail:yyhui2002@aliyun.com

基金项目:
国家自然科学基金项目(Nos.21904114,21765026)和云南省基础研究计划项目(Nos.202001AU070067,202201AT070028)资助。

摘要: 构建了一种基于BHQ-2和Cy5之间荧光共振能量转移(Fluorescence resonance energy transfer,FRET)的DNA纳米机器,用于前列腺特异性抗原(Prostate specific antigen,PSA)的检测。在燃料链(Fuel)的触发下,使整个DNA纳米机器产生熵增效应,引发FRET-DNA纳米机器循环运转,释放出大量的Cy5荧光信号探针,利用信号循环放大策略实现了对PSA的检测。通过琼脂糖凝胶电泳实验考察各步DNA反应杂交情况,优化了Fuel链浓度、孵育时间、适配体链浓度等实验条件。在最佳实验条件下,FRET-DNA纳米机器检测PSA的线性范围为0.1~100 ng/mL,检出限(3σ)低至93.3 pg/mL,与市售的传统PSA酶联免疫分析试剂盒比较,本方法表现出较宽的线性范围和较低的检出限。本方法具有无酶、低背景、高灵敏度、高选择性和操作方便等优势,并成功用于实际血清样品中PSA的检测,在生物医学检测领域具有良好的应用潜能。

关键词:

English

Fluorescence Resonance Energy Transfer-DNA Nanomachine-based Cycling Signal Amplified Strategy for Detection of Prostate Specific Antigen

College of Chemistry and Chemical Engineering, Yunnan Normal University, Kunming 650500, China

Corresponding author: YANG Tong, ; YANG Yun-Hui,

Received Date: 17 August 2021
Revised Date: 26 April 2022
Fund Project:
Supported by the National Natural Science Foundation of China (Nos.21904114, 21765026) and the Yunnan Fundamental Research Projects (Nos.202001AU070067, 202201AT070028).

Abstract: A fluorescence resonance energy transfer (FRET)-DNA nanomachine based on BHQ-2 and Cy5 was constructed for detection of prostate specific antigen (PSA). The introduction of fuel DNA would trigger the entropy increasing effect of the DNA nanomachine to cause the recycling of DNA nanomachine and release of more Cy5 signal probes, which further facilitated the cycling signal amplification for PSA analysis. The FRET-DNA nanomachine exhibited many advantages such as enzyme-free, low background, high sensitivity, high selectivity and easy operation. The DNA complementary hybridizations in working process of DNA nanomachine were characterized by agarose gel electrophoresis. Some important experimental parameters, such as the concentration and incubating time of fuel DNA, the concentration of PSA aptamer, were optimized. Under the optimized experimental conditions, the FRET-DNA nanomachine was employed to detect PSA protein with linearity ranging from 0.1 to 100 ng/mL and the limit of detection limit was 93.3 pg/mL (3σ). Comparing with commercial PSA kits based on enzyme-linked immunosorbent assay, this proposed strategy exhibited wider linearity rang and lower detection limit, which could be used to detect PSA in real serum samples. The easy operation and high reliability of this strategy showed potential quantitative power for future biomedical detection.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原

通讯作者: 杨通,E-mail:yangtong2018@ynnu.edu.cn; 杨云慧,E-mail:yyhui2002@aliyun.com

English

Fluorescence Resonance Energy Transfer-DNA Nanomachine-based Cycling Signal Amplified Strategy for Detection of Prostate Specific Antigen

Corresponding author: YANG Tong, ; YANG Yun-Hui,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原

通讯作者: 杨通,E-mail:yangtong2018@ynnu.edu.cn; 杨云慧,E-mail:yyhui2002@aliyun.com

English

Fluorescence Resonance Energy Transfer-DNA Nanomachine-based Cycling Signal Amplified Strategy for Detection of Prostate Specific Antigen

Corresponding author: YANG Tong, ; YANG Yun-Hui,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs