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Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization

机译:基于杂交链反应和酶诱导的金属化的级联扩增策略进行高灵敏的DNA检测

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摘要

A novel highly sensitive colorimetric assay for DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization was established. The DNA modified superparamagnetic beads were demonstrated to capture and enrich the target DNA in the hybridization buffer or human plasma. The hybridization chain reaction and enzyme-induced silver metallization on the gold nanoparticles were used as cascade signal amplification for the detection of target DNA. The metalization of silver on the gold nanoparticles induced a significant colour change from red to yellow until black depending on the concentration of the target DNA, which could be recognized by naked eyes. This method showed a good specificity for the target DNA detection, with the capabilty to discriminate single-base-pair mismatched DNA mutation (single nucleotide polymorphism). Meanwhile, this approach exhibited an excellent anti-interference capability with the convenience of the magentic seperation and washing, which enabled its usage in complex biological systems such as human blood plasma. As an added benefit, the utilization of hybridization chain reaction and enzyme-induced metallization improved detection sensitivity down to 10 pM, which is about 100-fold lower than that of traditional unamplified homogeneous assays.
机译:建立了一种新的高灵敏度比色测定法,用于基于杂交链反应和酶诱导的金属化的级联扩增策略的DNA检测。 DNA修饰的超顺磁珠被证明可以捕获并富集杂交缓冲液或人血浆中的目标DNA。金纳米颗粒上的杂交链反应和酶诱导的银金属化被用作级联信号扩增,以检测目标DNA。金纳米颗粒上银的金属化导致显着的颜色变化,从红色到黄色直至黑色,这取决于目标DNA的浓度,这可以通过肉眼识别。该方法对目标DNA检测显示出良好的特异性,并具有区分单碱基对错配DNA突变(单核苷酸多态性)的能力。同时,该方法显示出优异的抗干扰能力,并具有方便的分离和洗涤功能,从而使其可用于复杂的生物系统,例如人血浆。另外一个好处是,杂交链反应和酶诱导的金属化的利用将检测灵敏度降低至10 pM,这比传统的未扩增均相检测法低约100倍。

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