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Immobilization of CdSe/ZnS quantum dots on glass beads for the detection of nucleic acid hybridization using fluorescence resonance energy transfer

机译：将CdSe / ZnS量子点固定在玻璃珠上，以利用荧光共振能量转移检测核酸杂交

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The photoluminescence (PL) properties of quantum dots (QD) are of significant interest in the development of new methods for bioanalysis. Multiplexed solid-phase nucleic acid hybridization assays that use immobilized QDs as donors in fluorescence resonance energy transfer (FRET) are one such example, and offer several unique advantages over other methods. In this work, new interfacial chemistry is described for the immobilization of red-emitting CdSe/ZnS QDs on glass beads for use in hybridization assays. The beads were chemically modified with a dithiolate surface ligand and the QDs immobilized via self-assembly. Further derivatization of the QDs with dithiolate-terminated probe oligonucleotides enabled a hybridization assay that could detect unlabeled target down to nanomolar levels with discrimination of single base-pair mismatches. The use of beads as an immobilization platform afforded shorter analysis times and superior reusability compared to previous studies using optical fibers. Hybridization between probe, target, and Alexa Fluor 647 (A647) labeled reporter oligonucleotides in a sandwich format generated a spectroscopic signal by introducing the proximity needed for FRET between the QDs and A647. The results indicate clear directions for the optimization of solid-phase hybridization assays, and are important for the future development of true multiplexed biosensors based on QDs and FRET.
机译：量子点（QD）的光致发光（PL）特性在生物分析新方法的开发中引起了极大的兴趣。在荧光共振能量转移（FRET）中使用固定化QD作为供体的多重固相核酸杂交测定就是这样一个例子，与其他方法相比，它提供了许多独特的优势。在这项工作中，描述了一种新的界面化学，用于将红色发射的CdSe / ZnS QD固定在玻璃珠上，用于杂交测定。用二硫醇盐表面配体对珠进行化学修饰，并通过自组装固定QD。用二硫醇盐终止的探针寡核苷酸对QD进行进一步衍生化，可以进行杂交检测，该检测可以检测到低至纳摩尔水平的未标记靶标，并能识别单个碱基对错配。与以前使用光纤进行的研究相比，将珠子用作固定平台可以缩短分析时间，并具有更好的可重复使用性。探针，靶标和Alexa Fluor 647（A647）标记的报告寡核苷酸之间呈三明治形式的杂交通过在QD和A647之间引入FRET所需的邻近性而产生了光谱信号。结果为固相杂交测定的优化指明了明确的方向，对于基于QD和FRET的真正的多重生物传感器的未来开发非常重要。

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