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  

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

作　　者：李紫滢 李德燕 杨建梅[1] 胡蓉[1] 杨通 杨云慧[1] LI Zi-Ying;LI De-Yan;YANG Jian-Mei;HU Rong;YANG Tong;YANG Yun-Hui(College of Chemistry and Chemical Engineering,Yunnan Normal University,Kunming 650500,China)

机构地区：[1]云南师范大学化学化工学院,昆明650500

出　　处：《分析化学》2022年第7期1032-1040,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.21904114,21765026);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。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.

关 键 词：荧光共振能量转移 DNA纳米机器 循环信号放大 前列腺特异性抗原 生物医学检测 

分 类 号：R313[医药卫生—基础医学] O657.3[理学—分析化学]