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Hairpin Assembly Amplified Electrochemical Biosensor for Highly Sensitive and Specific Detection of DNA

机译：发夹组装扩增电化学生物传感器，用于DNA的高灵敏度和特异性检测

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In this report, a simple electrochemical biosensor has been developed for highly sensitive and specific detection of DNA based on hairpin assembly amplification. In the presence of target DNA, the biotin-labelled hairpin H1 is opened by hybridizing with target DNA through complementary sequences. Then the opened hairpin H1 assembles with the hairpin H2 to displace the target DNA, generating H1-H2 complex. The displaced target DNA could trigger the next cycle of hairpins assembly, resulting in the generation of numerous H1-H2 complexes. Subsequently, the H1-H2 complex hybridizes with the capture probe immobilized on the electrode. Fi-nally, the streptavidin alkaline phosphatase (ST-ALP) binds to biotin in the capture probe-H1-H2 complex and catalyzes the substrate alpha-naphthol (alpha-NP) to produce electrochemical signal. To make a more fascinating hairpin assembly amplification strategy in signal amplification, mismatched base sequences are designed in hairpin H2 to decrease non-specific binding of the hairpin substrates. The developed biosensor achieves a sensitivity of 20 pM with a linear range from 25 pM to 25 nM, and shows high selectivity toward single-base mismatch. Thus, the proposed electrochemical biosensor might have the potential for early clinical diagnosis and therapy.
机译：在此报告中，已经开发了一种简单的电化学生物传感器，用于基于发夹装配扩增对DNA进行高灵敏度和特异性检测。在目标DNA存在的情况下，通过互补序列与目标DNA杂交，可打开生物素标记的发夹H1。然后，打开的发夹H1与发夹H2组装在一起，以置换目标DNA，生成H1-H2复合物。置换的目标DNA可能触发发夹装配的下一个周期，从而导致产生大量H1-H2复合物。随后，H1-H2复合物与固定在电极上的捕获探针杂交。最后，链霉亲和素碱性磷酸酶（ST-ALP）与捕获探针-H1-H2复合物中的生物素结合，并催化底物α-萘酚（α-NP）以产生电化学信号。为了使信号放大中的发夹装配扩增策略更具吸引力，在发夹H2中设计了错配的碱基序列，以减少发夹底物的非特异性结合。研发的生物传感器在20 pM的线性范围从25 pM到25 nM的灵敏度范围内，对单碱基错配显示出高选择性。因此，提出的电化学生物传感器可能具有早期临床诊断和治疗的潜力。

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