Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya Suruga-Ku, Shizuoka, 422-8529, Japan.
Research Center for Vaccine and Drugs, National Research and Innovation Agency (BRIN), LAPTIAB 1, PUSPIPTEK, Tangerang Selatan, 15314, Indonesia.
Mikrochim Acta. 2024 Mar 4;191(4):174. doi: 10.1007/s00604-024-06259-7.
Early diagnosis of dengue infection by detecting the dengue virus non-structural protein 1 (DENV-NS1) is important to the patients to initiate speedy treatment. Enzyme-linked immunosorbent assay (ELISA)-based NS1 detection and RT-PCR are time-consuming and too complex to be employed in remote areas of dengue-endemic countries. Meanwhile, those of NS1 rapid test by lateral flow assay suffer from low detection limit. Electrochemical-based biosensors using screen-printed gold electrodes (SPGEs) have become a reliable detection method to convey both ELISA's high sensitivity and rapid test portability. In this research, we developed an electrochemical biosensor for DENV-NS1 detection by employing polydopamine (PDA)-modified SPGE. The electrodeposition of PDA on the surface of SPGE serves as a bioconjugation avenue for anti-NS1 antibody through a simple and low-cost immobilization procedure. The biosensor performance was evaluated to detect DENV-NS1 protein in PBS and human serum through a differential pulse voltammetric (DPV) technique. The developed sensing platform displayed a low limit of detection (LOD) of 1.63 pg mL and a wide linear range of 10 pg mL to 1 ng mL (R ∼ 0.969). The sensing platform also detected DEV-NS1 from four different serotypes in the clinical samples collected from dengue patients in India and Indonesia, with acceptable sensitivity, specificity, and accuracy values of 90.00%, 80.95%, and 87.65%, respectively. This result showcased the facile and versatile method of PDA coating onto the surface of screen-printed gold electrodes for a miniaturized point-of-care (PoC) detection device.
早期诊断登革热感染通过检测登革热病毒非结构蛋白 1（DENV-NS1）对患者及时进行治疗非常重要。基于酶联免疫吸附试验（ELISA）的 NS1 检测和 RT-PCR 耗时且过于复杂，无法在登革热流行国家的偏远地区使用。同时，侧向流动分析的 NS1 快速检测也存在检测限低的问题。基于电化学的生物传感器使用丝网印刷金电极（SPGE）已成为一种可靠的检测方法，兼具 ELISA 的高灵敏度和快速检测的便携性。在这项研究中，我们开发了一种基于电化学的生物传感器，用于通过使用聚多巴胺（PDA）修饰的 SPGE 检测 DENV-NS1。PDA 在 SPGE 表面的电沉积可作为通过简单且低成本的固定化程序将抗 NS1 抗体结合的途径。通过差分脉冲伏安法（DPV）技术，评估了该生物传感器在 PBS 和人血清中检测 DENV-NS1 蛋白的性能。所开发的传感平台在检测 1.63 pg mL 的低检测限（LOD）和 10 pg mL 至 1 ng mL 的宽线性范围（R ∼ 0.969）方面表现出色。该传感平台还从印度和印度尼西亚登革热患者采集的临床样本中检测到四种不同血清型的 DEV-NS1，其灵敏度、特异性和准确性分别为 90.00%、80.95%和 87.65%，具有可接受的值。该结果展示了将 PDA 简便且通用地涂覆到丝网印刷金电极表面的方法，可用于小型化即时检测（PoC）设备。
