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基于 PEI-CN/AuNWs 的电化学生物传感器用于通过外切酶辅助的信号放大测定氯霉素。

An electrochemical aptasensor based on PEI-CN/AuNWs for determination of chloramphenicol via exonuclease-assisted signal amplification.

机构信息

School of Food Science and Technology, Henan University of Technology, Zhengzhou High & New Technology Industries Development Zone, Lianhua Road 100#, Zhengzhou, 450001, Henan Province, People's Republic of China.

出版信息

Mikrochim Acta. 2021 Jan 6;188(1):22. doi: 10.1007/s00604-020-04688-8.

Abstract

An electrochemical aptasensor, including the polyethyleneimine-graphite-like carbon nitride/Au nanowire nanocomposite (PEI-CN/AuNWs) and exonuclease-assisted signal amplification strategy was constructed for the determination of chloramphenicol (CAP). Initially, a nanocomposite with substantial electrocatalytic property was synthesized by PEI-CN/AuNWs. This improves the conductivity and specific surface area of the PEI-CN/AuNW-modified gold electrode. Next, a DNA with a complementary sequence to a CAP aptamer (cDNA) was immobilized on the PEI-CN/AuNW-modified electrode, followed by the CAP aptamer hybridized with cDNA. The lower signal at this time is due to the negatively charged phosphate group of the oligonucleotide and [Fe (CN)] electrostatically repelling each other. The presence of the CAP would cause aptamer on the electrode surface to fall off and be digested by Recjf exonuclease, which resulted in target recycling, and a significant increase in DPV signal can be observed at a potential of 0.176 V (vs. Ag/AgCl). Under optimal conditions, there is a linear relationship between the peak current and the logarithm of CAP concentration in the range 100 fM-1 μM, and the detection limit of this aptasensor is 2.96 fM (S/N = 3). Furthermore, the resultant aptasensor has excellent specificity, reproducibility, and long-term stability, and has been applied to the detection of CAP in milk samples. Graphical abstract The detection principle of the electrochemical aptasensor for CAP detection was based on PEI-CN/AuNWs and exonuclease-assistant signal amplification. It is based on the fact that PEI-C3N4/AuNWs nanocomposites on the surface of the electrode can effectively improve the performance of the aptasensor, and Recjf exonuclease initiates the target recycling process, causes signal amplification.

摘要

一种电化学适体传感器,包括聚乙烯亚胺-类石墨相氮化碳/金纳米线纳米复合材料(PEI-CN/AuNWs)和外切酶辅助信号放大策略,用于检测氯霉素(CAP)。首先,通过 PEI-CN/AuNWs 合成具有大量电催化性能的纳米复合材料。这提高了 PEI-CN/AuNW 修饰金电极的导电性和比表面积。接下来,将与 CAP 适体互补序列的 DNA(cDNA)固定在 PEI-CN/AuNW 修饰的电极上,然后 CAP 适体与 cDNA 杂交。此时信号较低是由于寡核苷酸的带负电荷的磷酸基团和 [Fe(CN)] 之间的静电排斥。CAP 的存在会导致电极表面上的适体脱落并被 Recjf 外切酶消化,这导致了目标的循环利用,可以在 0.176 V(相对于 Ag/AgCl)的电位下观察到 DPV 信号的显著增加。在最佳条件下,在 100 fM-1 μM 的范围内,峰电流与 CAP 浓度的对数呈线性关系,该适体传感器的检测限为 2.96 fM(S/N=3)。此外,所得适体传感器具有优异的特异性、重现性和长期稳定性,并已应用于牛奶样品中 CAP 的检测。

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