Faculty of Science, La República University, Montevideo 11400, Uruguay.
Biosens Bioelectron. 2013 Mar 15;41:294-301. doi: 10.1016/j.bios.2012.08.044. Epub 2012 Sep 5.
Electrochemical DNA hybridization-based sensors show great promise as portable and automated analytical devices for routine screening of pathogenic or foreign nucleic acid sequences in biological samples. However, current sensor technologies still exhibit some unresolved issues which hampers their direct application into everyday life. Conducting polymers, such as polypyrrole (PPy), are increasingly being adopted as suitable platforms for DNA probe immobilization and signal transduction. Immobilization of DNA probes during pyrrole electropolymerization is a simple and efficient strategy to build composite electrodes suitable for DNA sensing. However, the effects of the probe state and sequence on PPy growth kinetics have not been studied yet. Here, we show that growth of PPy is drastically affected by the presence of guanine in the DNA probes and whether DNA is present in its single-stranded or double-stranded form. We show that some immobilization protocols may provoke irreversible oxidation of guanine moieties in the probe and that this issue deserves careful investigation as it may interfere with hybridization processes. We have also explored new procedures to build microelectrode arrays bearing immobilized DNA molecules, which are known to show beneficial properties in stirred samples. Overall, we present new techniques and concerns regarding the development of DNA-containing PPy-based composite electrodes, which may be taken into consideration for increasing genosensor reproducibility, response and performance.
电化学 DNA 杂交传感器作为一种便携式和自动化的分析设备,在生物样品中对病原体或外源核酸序列进行常规筛选方面具有广阔的应用前景。然而,当前的传感器技术仍存在一些尚未解决的问题,限制了其直接应用于日常生活。导电聚合物(如聚吡咯(PPy))越来越多地被用作 DNA 探针固定和信号转导的合适平台。在吡咯电聚合过程中固定 DNA 探针是构建适合 DNA 传感的复合电极的一种简单有效的策略。然而,探针状态和序列对 PPy 生长动力学的影响尚未得到研究。在这里,我们表明 PPy 的生长受到 DNA 探针中鸟嘌呤的存在以及 DNA 是处于单链还是双链形式的影响。我们表明,一些固定化方案可能会引起探针中鸟嘌呤部分的不可逆氧化,这是一个值得仔细研究的问题,因为它可能会干扰杂交过程。我们还探索了新的程序来构建带有固定化 DNA 分子的微电极阵列,这些分子在搅拌样品中显示出有益的性质。总的来说,我们提出了关于开发含 DNA 的 PPy 基复合电极的新技术和关注点,这些关注点可能会被考虑用于提高基因传感器的重现性、响应和性能。