Department of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran 1435116471, Iran.
Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran 1435116471, Iran.
Sensors (Basel). 2017 May 9;17(5):1074. doi: 10.3390/s17051074.
In this work, a novel nanocomposite consisting of electrosynthesized gold nanodendrites and chitosan nanoparticles (AuNDs/CSNPs) has been prepared to fabricate an impedimetric immunosensor based on a screen printed carbon electrode (SPCE) for the rapid and sensitive immunoassay of botulinum neurotoxin A (BoNT/A). BoNT/A polyclonal antibody was immobilized on the nanocomposite-modified SPCE for the signal amplification. The structure of the prepared nanocomposite was investigated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The charge transfer resistance (R) changes were used to detect BoNT/A as the specific immuno-interactions at the immunosensor surface that efficiently limited the electron transfer of Fe(CN)₆ as a redox probe at pH = 7.4. A linear relationship was observed between the %∆ and the concentration logarithm of BoNT/A within the range of 0.2 to 230 pg·mL with a detection limit (S/N = 3) of 0.15 pg·mL. The practical applicability of the proposed sensor was examined by evaluating the detection of BoNT/A in milk and serum samples with satisfactory recoveries. Therefore, the prepared immunosensor holds great promise for the fast, simple and sensitive detection of BoNT/A in various real samples.
在这项工作中,制备了一种由电合成的金纳米树突和壳聚糖纳米粒子(AuNDs/CSNPs)组成的新型纳米复合材料,用于基于丝网印刷碳电极(SPCE)的快速灵敏免疫传感器的制备,用于检测肉毒神经毒素 A(BoNT/A)。BoNT/A 多克隆抗体固定在纳米复合材料修饰的 SPCE 上以进行信号放大。通过透射电子显微镜(TEM)、扫描电子显微镜(SEM)、X 射线衍射(XRD)、傅里叶变换红外(FTIR)光谱、循环伏安法(CV)和电化学阻抗谱(EIS)研究了制备的纳米复合材料的结构。通过在 pH = 7.4 时在免疫传感器表面上发生的特定免疫相互作用来检测 BoNT/A,该相互作用有效地限制了作为氧化还原探针的 Fe(CN)₆的电子转移,从而改变了电荷转移电阻(R)变化。在 0.2 至 230 pg·mL 的范围内,观察到 %∆与 BoNT/A 浓度对数之间存在线性关系,检测限(S/N = 3)为 0.15 pg·mL。通过评估牛奶和血清样品中 BoNT/A 的检测,检验了所提出传感器的实际适用性,结果令人满意。因此,所制备的免疫传感器在各种实际样品中快速、简单、灵敏地检测 BoNT/A 具有很大的应用前景。