Department of Chemistry, University of Kurdistan, 66177-15175 Sanandaj, Iran.
Department of Chemistry, University of Kurdistan, 66177-15175 Sanandaj, Iran; Research Center for Nanotechnology, University of Kurdistan, 66177-15175 Sanandaj, Iran.
Biosens Bioelectron. 2018 Oct 15;117:332-339. doi: 10.1016/j.bios.2018.06.003. Epub 2018 Jun 4.
Development of simple, sensitive and specific method for human immunodeficiency virus (HIV) assays are urgently demand. In this study, we developed a novel molecularly imprinted polymer (MIP) electrochemiluminescence (MIP-ECL) sensor for the highly sensitive and selective HIV-1 gene detection using Europium sulfide nanocrystals (EsNCs) as signal producing compound. Here, the HIV aptamer as the template and o-phenylenediamine as the functional monomer, were electropolymerized directly on the surface of ITO electrode. With the hybridization reaction between the assemblies of EuS NCs functionalized 5-amino-labeled oligonucleotides as capture probes and oligonucleotides as detection target (HIV gene), the ECL signal significantly increased using KSO as coreactant. Taking advantage of both MIP-ECL assays and the strong electrochemiluminescence emission of EuS NCs, the sensitive and selective HIV gene detection has been achieved in a linear range 3.0 fM to 0.3 nM with a detection limit of 0.3 fM. The present MIP-ECL biosensor showed good specificity for HIV DNA detection compared to non-complementary and two bases mismatched sequences. The proposed ECL biosensor was applied to detect of HIV DNA in real human serum samples and satisfactory results were obtained. Due to high sensitivity and selectivity, excellent reproducibility and stability of the proposed assay, EuS NCs can be used as novel luminophore for development of MIP-ECL sensors for detection of other DNA biomarkers.
开发简单、灵敏和特异的人类免疫缺陷病毒 (HIV) 检测方法迫在眉睫。在这项研究中,我们开发了一种新型的分子印迹聚合物 (MIP) 电化学发光 (MIP-ECL) 传感器,用于高灵敏度和选择性的 HIV-1 基因检测,使用硫化铕纳米晶体 (EsNCs) 作为信号产生化合物。在这里,HIV 适体作为模板,邻苯二胺作为功能单体,直接在 ITO 电极表面电聚合。通过将功能化的 5-氨基标记寡核苷酸作为捕获探针的 EuS NCs 组装体与作为检测靶标 (HIV 基因) 的寡核苷酸进行杂交反应,使用 KSO 作为共反应物,ECL 信号显著增加。利用 MIP-ECL 分析和 EuS NCs 的强电化学发光发射,实现了在 3.0 fM 至 0.3 nM 的线性范围内对 HIV 基因的灵敏和选择性检测,检测限为 0.3 fM。与非互补和两个碱基错配序列相比,本研究开发的 MIP-ECL 生物传感器在 HIV DNA 检测方面具有良好的特异性。该 ECL 生物传感器已成功应用于人血清样本中 HIV DNA 的检测,获得了令人满意的结果。由于该检测方法具有高灵敏度、选择性、良好的重现性和稳定性,EuS NCs 可用作开发用于检测其他 DNA 生物标志物的 MIP-ECL 传感器的新型发光体。