Target-Catalyzed Dynamic Assembly-Based Pyrene Excimer Switching for Enzyme-Free Nucleic Acid Amplified Detection
Target-Catalyzed Dynamic Assembly-Based Pyrene Excimer Switching for Enzyme-Free Nucleic Acid Amplified Detection
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DOI：
10.1021/ac500834g
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发表时间：
2014-05-20
影响因子：
7.4
通讯作者：
He, Dinggeng
中科院分区：
文献类型：
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作者：
Qing, Zhihe;He, Xiaoxiao;He, Dinggeng
由于核酸作为生物靶标的内在重要性，核酸的简单和灵敏的检测对于生物学研究和医学诊断是非常必要的。本文将目标催化动态组装的信号放大能力与芘激基缔合物的空间灵敏荧光信号相结合，为无酶核酸扩增检测开辟了一条新的途径。在该策略中，设计了三个亚稳态芘标记的发夹DNA探针作为组装组分，其在靶DNA不存在的情况下在动力学上阻碍交叉打开。然而，在目标的存在下，分支结的动态组装被循环催化，并伴随着芘激基缔合物的开关，其在类似于488 nm处发射。因此，可以通过这种简单的混合检测扩增方法来检测靶DNA，而不需要昂贵且易腐烂的蛋白酶。该方法具有良好的检测能力，可检测的最低目标浓度为10 pM，与一些已报道的酶依赖性扩增方法相当甚至更好，验证了该方法在复杂流体中检测目标的潜力。此外，作为一种新的转换动态DNA组装技术到无酶信号放大分析应用，我们推断，所提出的策略将在更广泛的领域，包括适体为基础的非核酸靶传感，生物医学，生物成像具有广阔的应用前景。
Because of the intrinsic importance of nucleic acid as biotargets, the simple and sensitive detection of nucleic acid is very essential for biological studies and medical diagnostics. Herein, a new strategy for enzyme-free nucleic acid amplified detection has been opened up by combining the signal-amplification capability of target-catalyzed dynamic assembly with the spatially sensitive fluorescent signal of the pyrene excimer. In this strategy, three metastable pyrenelabeled hairpin DNA probes were designed as assembly components, which were kinetically handicapped from cross-opening in the absence, of the target DNA. However, in the presence of the target, the dynamic assembly of branched junctions was circularly catalyzed and accompanied by the switching of the pyrene excimer which emits at similar to 488 nm. Thus, the target DNA could be detected by this simple mix-and-detect amplification method, without expensive and perishable protein enzymes. A good detection capability exhibited with a detectable minimum target concentration of 10 pM, which was comparable to or even better than some reported enzyme-dependent amplification methods, and the potential for the target detection from complex fluids was verified. In addition, as a novel transformation of dynamic DNA assembly technology into enzyme-free signal-amplification analytical application, we infer that the proposed strategy will hold promising potential for application in a wider range of fields, including aptamer-based non-nucleic acid target sensing, biomedicine, and bioimaging.