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适体识别和邻近诱导熵驱动的无酶和快速扩增检测血小板衍生生长因子-BB 的电路。

Aptamer recognition and proximity-induced entropy-driven circuit for enzyme-free and rapid amplified detection of platelet-derived growth factor-BB.

机构信息

Department of Laboratory Medicine, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.

Department of Laboratory Medicine, Chongqing Traditional Chinese Medicine Hospital, Chongqing, Chongqing, 400016, China.

出版信息

Anal Chim Acta. 2019 Dec 27;1092:102-107. doi: 10.1016/j.aca.2019.09.046. Epub 2019 Sep 23.

DOI:10.1016/j.aca.2019.09.046
PMID:31708022
Abstract

Platelet-derived growth factor-BB (PDGF-BB) is currently used as a biomarker protein for cancer early diagnosis and clinical treatment. Herein, we reported a robust and enzyme-free strategy based on aptamer recognition and proximity-induced entropy-driven circuits (AR-PEDC) for homogeneous and rapid detection of platelet-derived growth factor BB (PDGF-BB) without any washing steps or thermocycling. The proximity probes specifically recognize target protein to form the completed trigger (CT). Then, the CT reacts with three-strand complex to form intermediate, which subsequently binds to fuel strand to release reporter strand, assistant strand and the CT. The revised proximity probes exhibit significantly improved signal-to-background ratio and faster association rate. Moreover, target protein/proximity probes interaction can specifically initiate entropy-driven circuits, thus providing immense signal amplification for ultrasensitive detection of PDGF-BB with low detection limit of 9.6 pM. The practical ability of the developed strategy is demonstrated by detection of PDGF-BB in human serum with satisfactory results. In addition, this method is flexible and can be conveniently extended to a variety of targets by simply substituting the target specific sequence. Thus, this strategy presents a rapid, low background and versatile amplification mechanism for the detection of protein biomarkers and offers a promising alternative platform for clinical diagnosis.

摘要

血小板衍生生长因子-BB(PDGF-BB)目前被用作癌症早期诊断和临床治疗的生物标志物蛋白。在此,我们报道了一种基于适体识别和近邻诱导熵驱动电路(AR-PEDC)的稳健且无需酶的策略,用于均相和快速检测血小板衍生生长因子 BB(PDGF-BB),无需任何洗涤步骤或热循环。近邻探针特异性识别靶蛋白以形成完整的触发(CT)。然后,CT 与三链复合物反应形成中间体,随后与燃料链结合释放报告链、辅助链和 CT。经修订的近邻探针表现出显著提高的信号与背景比和更快的缔合速率。此外,靶蛋白/近邻探针相互作用可以特异性引发熵驱动电路,从而为 PDGF-BB 的超灵敏检测提供巨大的信号放大,其检测限低至 9.6 pM。通过在人血清中检测 PDGF-BB 验证了所开发策略的实际能力,结果令人满意。此外,该方法具有灵活性,通过简单替换靶标特异性序列,即可方便地扩展到多种靶标。因此,该策略为蛋白质生物标志物的检测提供了一种快速、低背景和多功能的放大机制,并为临床诊断提供了一种有前途的替代平台。

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