Generic Neutravidin Biosensor for Simultaneous Multiplex Detection of MicroRNAs via Electrochemically Encoded Responsive Nanolabels
Generic Neutravidin Biosensor for Simultaneous Multiplex Detection of MicroRNAs via Electrochemically Encoded Responsive Nanolabels
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DOI：
10.1021/acssensors.8b00942
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发表时间：
2019-02-01
期刊：
影响因子：
8.9
通讯作者：
Mak, Wing Cheung
中科院分区：
文献类型：
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作者：
Azzouzi, Sawsen;Fredj, Zina;Mak, Wing Cheung
目前用于多种miRNA的电化学生物传感器需要繁琐地固定各种核酸探针。在这里，我们展示了一种创新的方法，使用一种通用的中性亲和素生物传感器结合电化学编码的响应纳米标记的miRNA-21和miRNA-141的简便和同时多重检测。生物传感器的选择性源于电化学编码的响应性纳米标记的固有性质，包括生物素化的分子信标（生物素-MB）和金属纳米颗粒（金属-NP）。该过程是简单的一锅法测定，其中靶向的miRNA引起生物素-MB的打开，随后通过中性亲和素生物传感器捕获生物素-MB-金属-NP，并通过溶出方波伏安法（SSWV）同时检测捕获的金属-NP。miRNA-21和miRNA-141的多重检测通过区分电化学特征（即，峰值电流）。该生物传感器同时检测miRNA-21的线性范围为0.5-1000 pM，检测限为0.3 pM（3 sigma/灵敏度，n = 3），miRNA-141的线性范围为50-1000 pM，检测限为10 pM。此外，我们证明了在加标血清样品中miRNA-21和miRNA-141的多重检测。
Current electrochemical biosensors for multiple miRNAs require tedious immobilization of various nucleic acid probes. Here, we demonstrate an innovative approach using a generic neutravidin biosensor combined with electrochemically encoded responsive nanolabels for facile and simultaneous multiplexed detection of miRNA-21 and miRNA-141. The selectivity of the biosensor arises from the intrinsic properties of the electrochemically encoded responsive nanolabels, comprising biotinylated molecular beacons (biotin-MB) and metal nanoparticles (metal-NPs). The procedure is a simple one-pot assay, where the targeted miRNA causes the opening of biotin-MB followed by capturing of the biotin-MB-metal-NPs by the neutravidin biosensor and simultaneous detection of the captured metal-NPs by stripping square-wave voltammetry (SSWV). The multiplexed detection of miRNA-21 and miRNA-141 is achieved by differentiation of the electrochemical signature (i.e., the peak current) for the different metal-NP labels. The biosensor delivers simultaneous detection of miRNAs with a linear range of 0.5-1000 pM for miRNA-21 and a limit of detection of 0.3 pM (3 sigma/sensitivity, n = 3), and a range of 50-1000 pM for miRNA-141, with a limit of detection of 10 pM. Furthermore, we demonstrate multiplexed detection of miRNA-21 and miRNA-141 in a spiked serum sample.