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Quantification of DNA by a Thermal-Durable Biosensor Modified with Conductive Poly(34-ethylenedioxythiophene)

机译:通过用导电聚(34-乙撑二氧噻吩)改性的热耐用生物传感器对DNA进行定量

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摘要

The general clinical procedure for viral DNA detection or gene mutation diagnosis following polymerase chain reaction (PCR) often involves gel electrophoresis and DNA sequencing, which is usually time-consuming. In this study, we have proposed a facile strategy to construct a DNA biosensor, in which the platinum electrode was modified with a dual-film of electrochemically synthesized poly(3,4-ethylenedioxythiophene) (PEDOT) resulting in immobilized gold nanoparticles, with the gold nanoparticles easily immobilized in a uniform distribution. The DNA probe labeled with a SH group was then assembled to the fabricated electrode and employed to capture the target DNA based on the complementary sequence. The hybridization efficiency was evaluated with differential pulse voltammetry (DPV) in the presence of daunorubicin hydrochloride. Our results demonstrated that the peak current in DPV exhibited a linear correlation the concentration of target DNA that was complementary to the probe DNA. Moreover, the electrode could be reused by heating denaturation and re-hybridization, which only brought slight signal decay. In addition, the addition of the oxidized form of nicotinamide adenine dinucleotide (NAD+) could dramatically enhance the sensitivity by more than 5.45-fold, and the limit-of-detection reached about 100 pM.
机译:在聚合酶链反应(PCR)之后进行病毒DNA检测或基因突变诊断的一般临床程序通常涉及凝胶电泳和DNA测序,这通常很耗时。在这项研究中,我们提出了一种构建DNA生物传感器的简便策略,其中用电化学合成的聚(3,4-乙撑二氧噻吩)(PEDOT)双层膜修饰铂电极,从而固定金纳米颗粒,金纳米粒子容易固定在均匀分布中。然后将标记有SH基团的DNA探针组装到装配好的电极上,并根据互补序列捕获目标DNA。在盐酸柔红霉素存在下,用差示脉冲伏安法(DPV)评估杂交效率。我们的结果表明,DPV中的峰值电流与与探针DNA互补的目标DNA浓度呈线性关系。而且,电极可以通过加热变性和再杂交来再利用,这只会带来轻微的信号衰减。另外,添加氧化型烟酰胺腺嘌呤二核苷酸(NAD + )可以显着提高灵敏度达5.45倍以上,检测限达到约100 pM。

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