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新型敏感电化学免疫传感器的开发用于选择性检测霍普氏菌生物标志物

Novel Sensitive Electrochemical Immunosensor Development for the Selective Detection of HopQ Bacteria Biomarker.

机构信息

Measurement and Sensor Technology, Chemnitz University of Technology, 09126 Chemnitz, Germany.

Department of Biomedical Engineering, Jordan University of Science and Technology, Irbid 22110, Jordan.

出版信息

Biosensors (Basel). 2023 May 8;13(5):527. doi: 10.3390/bios13050527.

Abstract

Helicobacter pylori () is a highly contagious pathogenic bacterium that can cause gastrointestinal ulcers and may gradually lead to gastric cancer. expresses the outer membrane HopQ protein at the earliest stages of infection. Therefore, HopQ is a highly reliable candidate as a biomarker for detection in saliva samples. In this work, an immunosensor is based on detecting HopQ as an biomarker in saliva. The immunosensor was developed by surface modification of screen-printed carbon electrodes (SPCE) with MWCNT-COOH decorated with gold nanoparticles (AuNP) followed by HopQ capture antibody grafting on SPCE/MWCNT/AuNP surface using EDC/S-NHS chemistry. The sensor performance was investigated utilizing various methods, such as cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscope (SEM) coupled with energy-dispersive X-ray spectroscopy (EDX). detection performance in spiked saliva samples was evaluated by square wave voltammetry (SWV). The sensor is suitable for HopQ detection with excellent sensitivity and linearity in the 10 pg/mL-100 ng/mL range, with a 2.0 pg/mL limit of detection (LOD) and an 8.6 pg/mL limit of quantification (LOQ). The sensor was tested in saliva at 10 ng/mL, and recovery of 107.6% was obtained by SWV. From Hill's model, the dissociation constant Kd for HopQ/HopQ antibody interaction is estimated to be 4.60 × 10 mg/mL. The fabricated platform shows high selectivity, good stability, reproducibility, and cost-effectiveness for early detection due to the proper choice of biomarker, the nanocomposite material utilization to boost the SPCE electrical performance, and the intrinsic selectivity of the antibody-antigen approach. Additionally, we provide insight into possible future aspects that researchers are recommended to focus on.

摘要

幽门螺杆菌()是一种高度传染性的致病菌,可导致胃肠道溃疡,并可能逐渐导致胃癌。在感染的早期阶段,表达外膜 HopQ 蛋白。因此,HopQ 是一种非常可靠的候选生物标志物,可用于唾液样本中的检测。在这项工作中,基于检测唾液中 HopQ 作为生物标志物的免疫传感器。该免疫传感器是通过在 MWCNT-COOH 上修饰金纳米粒子(AuNP)对丝网印刷碳电极(SPCE)进行表面修饰而开发的,然后通过 EDC/S-NHS 化学在 SPCE/MWCNT/AuNP 表面接枝 HopQ 捕获抗体。利用循环伏安法(CV)、电化学阻抗谱(EIS)和扫描电子显微镜(SEM)结合能谱(EDX)等多种方法研究了传感器的性能。通过方波伏安法(SWV)评估了在添加唾液样本中的检测性能。该传感器适用于 HopQ 检测,在 10 pg/mL-100 ng/mL 范围内具有优异的灵敏度和线性度,检测限(LOD)为 2.0 pg/mL,定量限(LOQ)为 8.6 pg/mL。该传感器在 10 ng/mL 的唾液中进行了测试,通过 SWV 获得了 107.6%的回收率。根据 Hill 模型,HopQ/HopQ 抗体相互作用的解离常数 Kd 估计为 4.60×10 mg/mL。由于选择了适当的生物标志物、利用纳米复合材料来提高 SPCE 的电性能以及抗体-抗原方法的固有选择性,该构建的平台具有高选择性、良好的稳定性、重现性和成本效益,可用于早期检测。此外,我们还提供了一些可能的未来研究方向的见解,建议研究人员关注这些方向。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58e9/10216805/f72df3d13555/biosensors-13-00527-g001.jpg

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