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基于目标触发的多通道 DNAzymes 循环扩增策略的电化学生物传感器,以 PtFe@Co-MOF 作为信号放大器。

An electrochemical aptasensor based on target triggered multiple-channel DNAzymes cycling amplification strategy with PtFe@Co-MOF as signal amplifier.

机构信息

School of Public Health, Chengdu Medical College, Chengdu, 610050, China.

Development and Regeneration Key Lab of Sichuan Province, Chengdu Medical College, Chengdu, 610050, China.

出版信息

Mikrochim Acta. 2022 Sep 21;189(10):388. doi: 10.1007/s00604-022-05478-0.

Abstract

A novel electrochemical aptasensor for the detection of Aflatoxin B1 (AFB1) was developed for the first time by using the target-triggered multiple-channel deoxyribozymes (DNAzymes) cycling amplified assay with Pt Fe doped NH-Co-MOF (PtFe@Co-MOF) as a signal amplifier. In the presence of AFB1, a self-assembling cross-over nucleic structure could be triggered by AFB1 via two aptamers' structure switching for strand displacement, resulting in four channels of Mg-dependent DNAzyme recycling simultaneously to multiply the detection signals. These DNAzymes cyclically split the substrate sequence to release the PtFe@Co-MOF labeled detection probe (DP), which is subsequently hybridized with the capture probes on the Au-deposited glassy carbon electrode. The fabrication procedure was characterized by differential pulse voltammetry, and the results of the morphological and element composition characteristics methods were analyzed to determine the successful preparation of PtFe@Co-MOF. The limit of detection (LOD) for AFB1 detection was 2 pg mL with a linear range from 5 pg mL to 80 ng mL. By comparison, the enhanced detection sensitivity has been found to originate from the efficient shearing of DNAzymes, enhanced peroxidase-like capability, and multiple active sites of PtFe@Co-MOF. Besides, this aptasensor showed high specificity for AFB1 compared with similar mycotoxins and exhibited high accuracy with low experimental cost and easy operation. Furthermore, the unique design of electrochemical aptasensors could provide a promising platform for the onsite determination of AFB1, as well as other targets by replacing the aptamer and other core recognition sequences.

摘要

一种新型电化学适体传感器,用于检测黄曲霉毒素 B1(AFB1),首次通过使用目标触发的多通道脱氧核酶(DNAzymes)循环放大测定法,利用 Pt Fe 掺杂 NH-Co-MOF(PtFe@Co-MOF)作为信号放大器来开发。在存在 AFB1 的情况下,通过两个适体的结构切换,通过 AFB1 触发自组装交叉核酸结构,用于链置换,从而同时实现四个通道的 Mg 依赖性 DNAzyme 循环再生,以倍增检测信号。这些 DNAzymes 循环分裂底物序列以释放 PtFe@Co-MOF 标记的检测探针(DP),随后与沉积金的玻碳电极上的捕获探针杂交。通过差分脉冲伏安法对制造过程进行了表征,并通过形态和元素组成特征方法的结果进行了分析,以确定 PtFe@Co-MOF 的成功制备。AFB1 检测的检测限(LOD)为 2pgmL,线性范围为 5pgmL 至 80ngmL。相比之下,已经发现增强的检测灵敏度源于 DNAzymes 的有效剪切、增强的过氧化物酶样能力以及 PtFe@Co-MOF 的多个活性位点。此外,与类似的真菌毒素相比,该适体传感器对 AFB1 表现出高特异性,并具有低实验成本和易于操作的高准确性。此外,电化学适体传感器的独特设计可以为 AFB1 以及其他目标的现场测定提供有前途的平台,方法是替换适体和其他核心识别序列。

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