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基于多纳米复合材料信号放大策略的降钙素原抗原电化学免疫分析。

Electrochemical immunoassay for procalcitonin antigen detection based on signal amplification strategy of multiple nanocomposites.

机构信息

School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510641, People's Republic of China.

出版信息

Biosens Bioelectron. 2014 Jan 15;51:310-6. doi: 10.1016/j.bios.2013.07.035. Epub 2013 Jul 26.

DOI:10.1016/j.bios.2013.07.035
PMID:23978454
Abstract

Procalcitonin, as a medium of inflammation, has become a new marker of the identification of severe bacterial infections in recent years and has received high attention due to its most ideal diagnostic indicators of specificity with major types of organism systemic inflammation of bacterial infection in the early stages. Thus, a novel method for the determination of procalcitonin (PCT) was developed based on a sandwich-type electrochemical immunosensor, which combined a simple immunosensor array as well as an effectively designed trace tag. The immunosensor was fabricated by layer-by-layer coating graphene (GC), carbon nanotubes (MWCNTs), chitosan (CS), glutaraldehyde (GA) composite on the working electrode, which can increase the electronic transfer rate and improve the surface area to capture a large number of primary antibodies (Ab1). The trace tag was prepared by loading high-content signal horseradish peroxidase labeled secondary PCT antibody (HRP-Ab2) with AuNPs, which were coated with mesoporous silica nanoparticles (MCM-41) through thionine linking. In comparison with conventional methods, the proposed immunosensor for PCT provided a better linear response range from 0.01 to 350 ng/mL and a lower limit of detection (LOD) of 0.5 pg/mL under optimal experimental conditions. In addition, the immunosensor exhibited convenience, low cost, rapidity, good specificity, acceptable stability and reproducibility. Moreover, satisfactory results were obtained for the determination of PCT in real human serum samples, indicating that the developed immunoassay has the potential to find application in clinical detection of PCT and other tumor markers as an alternative approach.

摘要

降钙素原作为炎症的中介物,近年来已成为严重细菌感染鉴别诊断的一个新标志物,因其在早期细菌感染全身炎症反应时,与主要病原体具有最佳的特异性诊断指标而受到高度关注。因此,基于三明治型电化学免疫传感器,开发了一种新的降钙素原(PCT)测定方法,该方法结合了简单的免疫传感器阵列和经过有效设计的痕量标记物。免疫传感器通过层层涂覆在工作电极上的石墨烯(GC)、碳纳米管(MWCNTs)、壳聚糖(CS)、戊二醛(GA)复合物来制备,这可以增加电子转移率并提高表面积,以捕获大量的初级抗体(Ab1)。痕量标记物通过将高含量信号辣根过氧化物酶标记的二级 PCT 抗体(HRP-Ab2)负载到金纳米颗粒(AuNPs)上,再通过硫堇键合涂覆介孔硅纳米颗粒(MCM-41)来制备。与传统方法相比,在最佳实验条件下,所提出的用于 PCT 的免疫传感器提供了更好的线性响应范围(从 0.01 到 350ng/mL)和更低的检测限(LOD)(0.5pg/mL)。此外,该免疫传感器具有方便、低成本、快速、良好的特异性、可接受的稳定性和重现性。此外,该免疫传感器在实际人血清样本中测定 PCT 时取得了令人满意的结果,表明所开发的免疫分析具有作为替代方法在临床检测 PCT 和其他肿瘤标志物中的应用潜力。

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