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一种基于丝网印刷碳电极的用于检测人诺如病毒的宽范围一次性电化学生物传感器。

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

作者信息

Wang Nan, Pan Guiying, Guan Shimin, Rong Shaofeng, Wang Dapeng, Gao Zhiyong, Tian Peng, Li Qianqian

机构信息

Department of Bioengineering, Shanghai Institute of Technology, Shanghai, China.

Department of Food Science and Technology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.

出版信息

Front Bioeng Biotechnol. 2022 Mar 18;10:845660. doi: 10.3389/fbioe.2022.845660. eCollection 2022.

Abstract

Human noroviruses (HuNoVs) are the major non-bacterial pathogens causing acute gastroenteritis in people of all ages worldwide. No stable culture system 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 (≈1.93×10 HuNoVs/mL) and 0.22 ng/ml (≈1.15×10 HuNoVs/mL), respectively. The detection limitation of both GI and GII HuNoVs in clinical fecal samples was 10 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.

摘要

人诺如病毒(HuNoVs)是全球各年龄段人群急性胃肠炎的主要非细菌性病原体。目前尚无稳定的培养系统用于常规检测多种HuNoV毒株。一种简单快速的HuNoV检测方法对于防控这种病原体具有重要意义。在本研究中,基于丝网印刷碳电极(SPCE)构建了一种用于灵敏快速检测HuNoV的电化学生物传感器。金纳米颗粒和蛋白A分别应用于SPCE表面以增强电信号和使抗体以固定方向连接。进一步将针对病毒衣壳(VP1)S结构域蛋白的单克隆抗体(MAb)固定在SPCE上以特异性结合HuNoV。VP1与包被的MAb结合导致通过循环伏安法(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/HuNoV的浓度相关。基因群I.1(GI.1)VP1和基因群II.4(GII.4)VP1的检测限分别为0.37 ng/ml(≈1.93×10 HuNoV/mL)和0.22 ng/ml(≈1.15×10 HuNoV/mL)。临床粪便样本中GI和GII HuNoV的检测限均为10个基因组拷贝/mL。结果可在1小时内获得。我们证明这种一次性电化学生物传感器是快速检测不同基因群和基因型HuNoV的良好候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f5ad/8992780/2edb097029fa/fbioe-10-845660-g001.jpg

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