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利用辣根过氧化物酶模拟 DNA 酶作为适体传感器检测乳腺癌来源的外泌体。

Detection of breast cancer-derived exosomes using the horseradish peroxidase-mimicking DNAzyme as an aptasensor.

机构信息

Lab of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

出版信息

Analyst. 2019 Dec 16;145(1):107-114. doi: 10.1039/c9an01653h.

Abstract

Exosomes are membrane-enclosed phospholipid extracellular vesicles with a variety of tumor antigens which can be applied in the diagnosis and treatment of cancer due to the high secretion on the surface of cancer cells. Until now, many research studies on exosomes have been reported, but convenient and low-cost detection methods still need to be developed. Recently, we have developed a sensitive, simple and low-cost colorimetric aptasensor by designing a hairpin-like structure, which combined the highly specific MUC1 aptamer with a hemin/G-quadruplex for the detection of breast cancer exosomes. The hemin/G-quadruplex toward H2O2 reduction was used to generate an evidently strong colorimetric response owing to acting as a HRP-mimicking DNAzyme. The aptasensor is regarded as an "on-off" type switch, which strictly controls the process of reaction responding to the existence or not of exosomes. In our study, associated exosome detection limits are 3.94 × 105 particles per mL, which showed a higher sensitivity compared to commercial ELISA. Our method not only exhibited the advantages of being convenient and time-saving, with few instruments used, but also the signal generated using this method can be easily observed by the naked eye. Furthermore, the proposed strategy can well differentiate breast cancer patients from healthy individuals, demonstrating potential application in the analysis of clinical specimens.

摘要

外泌体是一种带有多种肿瘤抗原的膜封闭磷脂双层细胞外囊泡,由于其在癌细胞表面的高分泌,可应用于癌症的诊断和治疗。到目前为止,已经有许多关于外泌体的研究报告,但仍需要开发方便且低成本的检测方法。最近,我们通过设计发夹状结构,开发了一种灵敏、简单且低成本的比色适体传感器,该结构将高度特异性的 MUC1 适体与血红素/G-四链体结合,用于检测乳腺癌外泌体。由于血红素/G-四链体可作为 HRP 模拟 DNA 酶还原 H2O2,因此可产生明显强烈的比色响应。该适体传感器被视为“开-关”型开关,严格控制反应过程,以响应外泌体的存在或不存在。在我们的研究中,相关外泌体检测下限为 3.94×105 个/毫升,与商业 ELISA 相比具有更高的灵敏度。我们的方法不仅具有方便、省时、使用仪器少的优点,而且该方法产生的信号可以用肉眼轻松观察到。此外,该策略可以很好地区分乳腺癌患者和健康个体,证明了其在临床标本分析中的潜在应用。

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