Simultaneous detection of microcysin-LR and okadaic acid using a dual fluorescence resonance energy transfer aptasensor
Simultaneous detection of microcysin-LR and okadaic acid using a dual fluorescence resonance energy transfer aptasensor
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使用双荧光共振能量转移适体传感器同时检测小藻毒素-LR 和冈田酸
DOI：
10.1007/s00216-014-8378-3
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发表时间：
2015
影响因子：
4.3
通讯作者：
Wang Zhouping
中科院分区：
文献类型：
--
作者：
Wu Shijia;Duan Nuo;Zhang Hui;Wang Zhouping
藻类毒素可通过食用受污染的水和食物引起人类的神经毒性、肝毒性和细胞毒性。本研究提出了一种新的适配子，用于同时检测微囊藻毒素-LR（MC-LR）和冈田酸（OA）。该体系采用绿色和红色上转换纳米粒子（UCNP）发光作为供体，两种猝灭剂（BHQ 1和BHQ 3）作为相应的受体。通过将适体与其相应的互补DNA杂交来制造两个供体-受体对。结果表明，由于荧光光谱的高度重叠，猝灭剂探针可以猝灭绿色和红色上转换发光。在MC-LR和OA的存在下，适体优选与其相应的分析物结合并与互补DNA去杂交。该效应变得足够大以防止绿色和红色发光猝灭。在优化的实验条件下，随着藻毒素浓度的增加，相对发光强度增加，允许定量的MC-LR和OA。MC-LR和OA在0.1 ~ 50 ng mL− 1范围内发光强度与藻毒素浓度的曲线图呈线性关系。作为一个实际应用，这种类型的双荧光共振能量转移（FRET）适体传感器被用来监测自然污染的食品样品，如鱼和虾中的MC-LR和OA水平。图示摘要双FRET适体传感器之间的上转换纳米粒子和猝灭剂的示意图，用于同时检测MC-LR和OA
Algal toxins can cause neurovirulence, hepatotoxicity, and cytotoxicity in humans through the consumption of contaminated water and food. In this work, we presented a novel aptasensor for the simultaneous detection of two algal toxins, microcysin-LR (MC-LR) and okadaic acid (OA). This system employed green and red upconversion nanoparticle (UCNP) luminescence as the donors and two quenchers (BHQ1and BHQ3) as the corresponding acceptors. The two donor–acceptor couples were fabricated by hybridizing the aptamers with their corresponding complementary DNA. The results indicated that the green and red upconversion luminescence could be quenched by the quencher probes because of their highly overlapping spectrum. In the presence of MC-LR and OA, the aptamers preferred to bind to their corresponding analytes and de-hybridize with the complementary DNA. This effect became sufficiently large to prevent green and red luminescence quenching. Under the optimized experimental conditions, the relative luminescence intensity increased as the algal toxin concentrations increased, allowing for the quantification of MC-LR and OA. The relationships between the luminescence intensity and plotting logarithms of algal toxin concentrations were linear in the range from 0.1 to 50 ng mL−1for MC-LR and OA. As a practical application, this type of dual fluorescence resonance energy transfer (FRET) aptasensor was used to monitor the MC-LR and OA levels in naturally contaminated food samples such as fish and shrimps.Graphical AbstractSchematic illustration of dual FRET aptasensor between the upconversion nanoparticles and quenching agent for the simultaneous detection of MC-LR and OA