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首页> 外文期刊>Biosensors & Bioelectronics: The International Journal for the Professional Involved with Research, Technology and Applications of Biosensers and Related Devices >Label-free chemiluminescent aptasensor for platelet-derived growth factor detection based on exonuclease-assisted cascade autocatalytic recycling amplification
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Label-free chemiluminescent aptasensor for platelet-derived growth factor detection based on exonuclease-assisted cascade autocatalytic recycling amplification

机译：基于核酸外切酶辅助级联自催化循环扩增的无标记化学发光适体传感器，用于血小板源性生长因子检测

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摘要

Here an exonuclease III (Exo III)-assisted cascade autocatalytic recycling amplification (Exo-CARA) strategy is proposed for label-free chemiluminescent (CL) detection of platelet-derived growth factor BB (PDGF-BB) by taking advantage of both recognition property of aptamer and cleavage function of Exo III. Functionally, this system consists of a duplex DNA (aptamer-blocker hybrid), two kinds of hairpin structures (MB_1 and MB_2), and Exo III. Upon recognizing and binding with PDGF-BB, aptamer folds into a close configuration, which initiates the proposed Exo-CARA reaction (Recyclings I→II→III→II). Finally, numerous "caged" G-quadruplex sequences on DNAzyme_1 and DNAzyme_2 release that intercalate hemin to catalyze the oxidation of luminol by H_2O_2 to generate an amplified CL signal, achieving excellent specificity and high sensitivity with a detection limit of 6.8 ×10~(-13) M PDGF-BB. The proposed strategy has the advantages of simple design, isothermal conditions, homogeneous reaction without separation and washing steps, effective-cost without the need of labeling, and high amplification efficiency, which might be a universal and promising protocol for the detection of a variety of biomolecules whose aptamers undergo similar conformational changes.
机译：本文提出了一种核酸外切酶III（Exo III）辅助级联自动催化循环扩增（Exo-CARA）策略，通过利用两者的识别特性来对血小板衍生的生长因子BB（PDGF-BB）进行无标记化学发光（CL）检测。 Exo III的适体和裂解功能从功能上讲，该系统由双链体DNA（适体-阻断剂杂交体），两种发夹结构（MB_1和MB_2）和Exo III组成。识别并与PDGF-BB结合后，适体折叠成紧密构型，从而引发拟议的Exo-CARA反应（循环I→II→III→II）。最后，DNAzyme_1和DNAzyme_2上释放出许多“笼中的” G-四链体序列，这些序列嵌入血红素中，以催化H_2O_2氧化鲁米诺，从而产生放大的CL信号，从而实现了出色的特异性和高灵敏度，检测限为6.8×10〜（- 13）M PDGF-BB。所提出的策略具有设计简单，等温条件，无需分离和洗涤步骤即可进行均相反应，无需标记的有效成本以及扩增效率高的优点，这可能是检测多种不同形式的通用且有希望的方案适体经历相似构象变化的生物分子。

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