DataSheet1_A Broad-Range Disposable Electrochemical Biosensor Based on Screen-Printed Carbon Electrodes for Detection of Human Noroviruses.PDF

Human noroviruses (HuNoVs) are the major non-bacterial pathogens causing acute gastroenteritis in people of all ages worldwide. No stable culture system in vitro is available for routing the detection of multiple strains of HuNoVs. A simple and rapid method for detection of HuNoVs is of great significance for preventing and controlling this pathogen. In this work, an electrochemical biosensor for sensitive and fast detection of HuNoVs was constructed based on a screen-printed carbon electrode (SPCE). Gold nanoparticles and protein-A were applied on the SPCE surface for enhancement of the electrical signals and the linkage of antibodies with a fixed orientation, respectively. A monoclonal antibody (MAb) against the S domain protein of the viral capsid (VP1) was further immobilized on the SPCE to bind HuNoVs specifically. The binding of VP1 to the coated MAbs resulted in the reduction of conductivity (current) measured by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The reduction in the current was correlated to the concentration of VP1/HuNoVs. The detection limitation of Genogroup I.1 (GI.1) VP1 and Genogroup II.4 (GII.4) VP1 was 0.37 ng/ml (≈1.93×107 HuNoVs/mL) and 0.22 ng/ml (≈1.15×107 HuNoVs/mL), respectively. The detection limitation of both GI and GII HuNoVs in clinical fecal samples was 104 genomic copies/mL. The results could be obtained in 1 h. We demonstrated that this disposable electrochemical biosensor was a good candidate for rapid detection of different genogroup and genotype HuNoVs.

人类诺如病毒（HuNoVs）是全球范围内引起各年龄层人群急性胃肠炎的主要非细菌性病原体。目前尚无稳定的体外培养系统用于HuNoVs多个菌株的常规检测。因此，开发一种简单快速检测HuNoVs的方法对于预防和控制该病原体具有重要意义。在本研究中，基于丝网印刷碳电极（SPCE）构建了一种用于敏感和快速检测HuNoVs的电化学生物传感器。金纳米粒子和蛋白A被应用于SPCE表面，以增强电信号和实现抗体与固定方向的连接。进一步地，将针对病毒衣壳S蛋白（VP1）的单克隆抗体（MAb）固定于SPCE上，以特异性结合HuNoVs。VP1与涂覆的MAbs的结合导致循环伏安法（CV）和差分脉冲伏安法（DPV）测量的电导率（电流）降低，电流的降低与VP1/HuNoVs的浓度相关。Genogroup I.1（GI.1）VP1和Genogroup II.4（GII.4）VP1的检测限分别为0.37 ng/ml（≈1.93×10^7 HuNoVs/mL）和0.22 ng/ml（≈1.15×10^7 HuNoVs/mL），而GI和GII HuNoVs在临床粪便样本中的检测限为104基因组拷贝/mL。检测结果可在1小时内获得。我们证明了这种一次性电化学生物传感器是快速检测不同基因组和基因型HuNoVs的良好候选者。