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一锅法制备二氨基萘 -AuNPs 修饰的石墨烯纳米平台,用于生物样品中耐甲氧西林金黄色葡萄球菌的检测。

One pot fabrication of diamino naphthalene -AuNPs decorated graphene nanoplatform for the MRSA detection in the biological sample.

机构信息

Department of Pharmaceutical Chemistry, College of Health Sciences, University of KwaZulu-Natal, Westville Campus, Durban 4000, South Africa.

Department of Pharmaceutical Chemistry, College of Health Sciences, University of KwaZulu-Natal, Westville Campus, Durban 4000, South Africa.

出版信息

Bioelectrochemistry. 2024 Jun;157:108674. doi: 10.1016/j.bioelechem.2024.108674. Epub 2024 Feb 28.

DOI:10.1016/j.bioelechem.2024.108674
PMID:38460467
Abstract

Early monitoring of MRSA can effectively mitigate the disease risk by using Penicillin-binding protein 2a (PbP2a) biomarker. Diamino naphthalene-AuNPs decorated graphene (AuNPsGO-DN) nanocomposite was synthesized for a rapid and sensitive immunosensor detecting PbP2a. The synthesized AuNPsGO-DN nanocomposites were characterized by field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (SEM-EDX), Fourier transform infrared spectroscopy (FT-IR), Raman spectroscopy, and X-ray diffraction spectroscopy (XRD). Electrochemical characterization done with cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrical impedance spectroscopy (EIS) techniques. Anti-PbP2a monoclonal antibodies immobilized at AuNPsGO-DN/GCE via covalent bonding. AuNPs enhanced the electrode surface area and the antibodies' loading. Mercaptopropionic acid (MPA) was a linker between the AuNPs and antibodies, orientated the antibodies as opposite to the PbP2a antigen, and improved the sensitivity and specificity. The antiPbP2a/MPA/AuNPsGO-DN/GCE electrode displayed sensitive and selective detection towards the PbP2a antigen in phosphate buffer saline (PBS pH 7.4). The broad linear range from 0.01 to 8000 pg/mL was obtained with LOD of 0.154 pg/mL and 0.0239 pg/mL, respectively. A label-free, simple, and sensitive immunosensor was developed with a 98-106 % recovery rate in spiked biological samples. It shows the potential applicability of the developed immunoelectrode.

摘要

早期监测耐甲氧西林金黄色葡萄球菌(MRSA)可以通过使用青霉素结合蛋白 2a(PbP2a)生物标志物有效降低疾病风险。本研究合成了二氨基萘-AuNPs 修饰的石墨烯(AuNPsGO-DN)纳米复合材料,用于快速灵敏地检测 PbP2a 的免疫传感器。通过场发射扫描电子显微镜(FE-SEM)、能量色散 X 射线光谱(SEM-EDX)、傅里叶变换红外光谱(FT-IR)、拉曼光谱和 X 射线衍射光谱(XRD)对合成的 AuNPsGO-DN 纳米复合材料进行了表征。通过循环伏安法(CV)、差分脉冲伏安法(DPV)和电化学阻抗谱(EIS)技术进行电化学表征。通过共价键将抗 PbP2a 单克隆抗体固定在 AuNPsGO-DN/GCE 上。AuNPs 增加了电极表面积和抗体的负载量。巯基丙酸(MPA)是 AuNPs 和抗体之间的连接体,使抗体与 PbP2a 抗原相对定向,并提高了灵敏度和特异性。抗 PbP2a/MPA/AuNPsGO-DN/GCE 电极在磷酸盐缓冲盐水(PBS pH 7.4)中对 PbP2a 抗原表现出敏感和选择性的检测。在 0.01 至 8000 pg/mL 的宽线性范围内,检测限分别为 0.154 pg/mL 和 0.0239 pg/mL。开发了一种无标记、简单、灵敏的免疫传感器,在生物样品中加标回收率为 98-106%。它显示了所开发免疫电极的潜在适用性。

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