Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.
Colloids Surf B Biointerfaces. 2013 Dec 1;112:344-9. doi: 10.1016/j.colsurfb.2013.07.027. Epub 2013 Jul 24.
An efficient DNA electrochemical biosensor, based on the gold nanoparticles (GNPs) in situ synthesized at the surface of multiwalled carbon nanotubes (MWCNTs), cerium dioxide (CeO2) and chitosan (Chits) composite membrane, was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). The capture probe was attached onto the nanocomposite membrane modified glassy carbon electrode (GCE) through the conjugated structure. Owing to the synergistic effects of CeO2 nanoparticles with a strong adsorption ability and MWCNTs with a large surface area and excellent electron transfer ability, the prepared composite membrane was demonstrated an efficient electron transfer ability. The biosensor was electrochemically characterized by cyclic voltammogram (CV) and differential pulse voltammetry (DPV), and the decrease of the peak currents upon hybridization was observed using methylene blue (MB) as the electroactive indicator. Under the optimized conditions, peak currents were linear over the range from 1 × 10(-9) M to 1 × 10(-)(12) M, with a detection limit of 5 × 10(-)(13) M (based on the 3σ). And the proposed method was successfully applied for the detection of PCR real samples with satisfactory results. Furthermore, the developed DNA biosensor was demonstrated a good selectivity, a reasonable stability and a favorable reproducibility, which could be regenerated easily.
一种基于金纳米粒子(GNPs)原位合成的多壁碳纳米管(MWCNTs)、二氧化铈(CeO2)和壳聚糖(Chits)复合膜的高效 DNA 电化学生物传感器,被开发用于检测慢性髓性白血病(CML)中的 BCR/ABL 融合基因。捕获探针通过共轭结构附着在纳米复合膜修饰的玻碳电极(GCE)上。由于 CeO2 纳米粒子具有很强的吸附能力,MWCNTs 具有较大的表面积和优异的电子传递能力,因此制备的复合膜具有高效的电子传递能力。通过循环伏安法(CV)和差分脉冲伏安法(DPV)对生物传感器进行电化学表征,并观察到杂交后亚甲蓝(MB)作为电活性指示剂的峰电流减小。在优化条件下,峰电流在 1×10(-9)M 至 1×10(-12)M 的范围内呈线性关系,检测限为 5×10(-13)M(基于 3σ)。该方法成功应用于 PCR 实际样品的检测,结果令人满意。此外,所开发的 DNA 生物传感器具有良好的选择性、合理的稳定性和良好的重现性,且易于再生。