Sequence-specific, electronic detection of oligonucleotides in blood, soil, and foodstuffs with the reagentless, reusable E-DNA sensor
Sequence-specific, electronic detection of oligonucleotides in blood, soil, and foodstuffs with the reagentless, reusable E-DNA sensor
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DOI：
10.1021/ac0601819
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发表时间：
2006-08-15
影响因子：
7.4
通讯作者：
Plaxco, Kevin W.
中科院分区：
文献类型：
--
作者：
Lubin, Arica A.;Lai, Rebecca Y.;Plaxco, Kevin W.
在不使用外源性试剂和使用可重复使用的全电子平台的情况下，检测复杂的污染物污染样品中的特定寡核苷酸的能力可以彻底改变临床和现场的病原体检测。在这里，我们描述了一种无标记的电子传感器，称为E-DNA，它能够同时满足这些具有挑战性的需求。我们发现，由于信号的产生是耦合到一个杂交连接的构象变化，而不是只吸附到传感器表面，E-DNA是有足够的选择性来检测复杂的，多组分的样品，如血清和土壤中的寡核苷酸。此外，E-DNA信号与靶互补性单调相关，允许传感器区分错配的靶：我们很容易检测到互补的17个碱基的靶对50 000倍过量的基因组DNA，可以直接在血清中区分三个碱基的错配与完美的靶，并且在理想条件下，观察到单碱基错配之间的统计学显著差异。最后，由于传感元件连接到电极表面，E-DNA是可重复使用的：30秒的室温洗涤可恢复> 99%的传感器信号。这项工作进一步支持了E-DNA作为一种快速，特异，方便的DNA和RNA序列检测方法的实用性。
The ability to detect specific oligonucleotides in complex, contaminant-ridden samples, without the use of exogenous reagents and using a reusable, fully electronic platform could revolutionize the detection of pathogens in the clinic and in the field. Here, we characterize a label-free, electronic sensor, termed E-DNA, for its ability to simultaneously meet these challenging demands. We find that because signal generation is coupled to a hybridization-linked conformational change, rather than to only adsorption to the sensor surface, E-DNA is selective enough to detect oligonucleotides in complex, multicomponent samples, such as blood serum and soil. Moreover, E-DNA signaling is monotonically related to target complementarity, allowing the sensor to discriminate between mismatched targets: we readily detect the complementary 17-base target against a 50 000-fold excess of genomic DNA, can distinguish a three-base mismatch from perfect target directly in blood serum, and under ideal conditions, observe statistically significant differences between single-base mismatches. Finally, because the sensing components are linked to the electrode surface, E-DNA is reusable: a 30-s room temperature wash recovers > 99% of the sensor signal. This work further supports the utility of E-DNA as a rapid, specific, and convenient method for the detection of DNA and RNA sequences.