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Electrochemical aptasensing strategy for kanamycin detection based on target-triggered single-strand DNA adsorption on MoS_2 nanosheets and enzymatic signal amplification

机译:基于目标触发单链DNA吸附对MOS_2纳米蛋白和酶联酶扩增的Kanamycin检测电化学Aptasening策略

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A selective and sensitive electrochemical aptasensing strategy was developed for kanamycin detection based on target-triggered single-strand DNA adsorption on MoS2 nanosheets and enzymatic signal amplification. Firstly, a Y-shape double-stranded DNA (dsDNA) was designed containing probe DNA, assist DNA (labeled with biotin at its 3'-terminal) and aptamer DNA. Then, in the presence of kanamycin, aptamer DNA can be released from the Y-shape dsDNA structure due to its interaction with kanamycin to form stable conjugate, which will destroy the Y-shape dsDNA structure and lead to the presence of two-fragment of single-stranded DNA (ssDNA) on the remained half-hybrid DNA structure (containing probe DNA and assist DNA). Based on the strong interaction between ssDNA and MoS2 nanosheets, the half-hybrid DNA structure can be captured on MoS2 nanosheets modified electrode surface, which triggers the subsequent immobilization of streptavidin-alkaline phosphatase (SA-ALP) compound. Under the catalysis of ALP, p-nitrophenol phosphate (PNPP) is hydrolyzed to produce p-nitrophenol (PNP). Based on the linear relationship between the electrochemical oxidation signal of PNP and kanamycin concentration, kanamycin can be detected with the detection limit of 0.029 nM (S/N = 3). The aptamer-MoS2 nanosheets based electrochemical biosensor shows good selectivity, reproducibility for kanamycin detection.
机译:基于目标触发的单链DNA吸附和酶促信号扩增,为基于目标触发的单链DNA吸附而开发了一种选择性和敏感的电化学Aptasensing策略。首先,设计含有探针DNA的Y形双链DNA(DSDNA),辅助DNA(在其3'-末端的生物素标记)和适体DNA。然后,在Kanamycin的存在下,由于其与卡那霉素的相互作用以形成稳定的缀合物,可以从Y形DSDNA结构中释放Aptamer DNA,这将破坏Y形DSDNA结构并导致两种片段的存在在剩余的半杂交DNA结构上(含有探针DNA和辅助DNA)上的单链DNA(SSDNA)。基于SSDNA和MOS2纳米片之间的强相互作用,可以在MOS2纳米蛋白酶改性电极表面上捕获半杂化DNA结构,其触发了链霉抗生物素蛋白 - 碱性磷酸酶(SA-ALP)化合物的随后固定。在AlP的催化下,水解p-硝基苯酚磷酸盐(PNPP)以产生p-硝基苯酚(PNP)。基于PNP和卡那霉素浓度的电化学氧化信号之间的线性关系,通过0.029nm的检测限/ / n = 3)可以检测卡那霉素。基于Aptamer-MOS2纳米片的电化学生物传感器显示出良好的选择性,用于卡那霉素检测的再现性。

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