Simple approach for ultrasensitive electrochemical immunoassay of Clostridium difficile toxin B detection
Simple approach for ultrasensitive electrochemical immunoassay of Clostridium difficile toxin B detection
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艰难梭菌毒素 B 检测的超灵敏电化学免疫分析的简单方法
DOI：
10.1016/j.bios.2013.09.063
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发表时间：
2014-03-15
影响因子：
12.6
通讯作者：
Wang, Ju-Fang
中科院分区：
文献类型：
--
作者：
Fang, Yi-Shan;Chen, Shu-Yi;Wang, Ju-Fang
艰难梭菌毒素B（Clostridium difficile toxin，Tcd B）是艰难梭菌相关性疾病的主要致病因子之一，因其毒力强、产孢能力强、持续性强等特点，已成为临床诊断的潜在生物标志物。因此，基于三明治型电化学免疫传感器，开发了一种简单的测定Tcd B的方法。通过将碳纳米管（MWCNTs）、普鲁士蓝（PB）、壳聚糖（CS）、戊二醛（GA）复合物逐层涂覆在工作电极上，并以氧化石墨烯（GO）为载体，采用多酶放大策略，制备了免疫传感器，实现了对多酶的灵敏度大幅提高。与传统方法相比，该方法具有较高的灵敏度和选择性，在最佳实验条件下，Tcd B的线性范围为0.003 ~ 320 ng/mL，检测下限为0.7 pg/mL（S/N=3）。该免疫传感器具有方便、廉价、快速、特异性好、稳定性和重现性好等优点。该方法用于人粪便标本中Tcd B的检测，结果令人满意，表明该方法可作为Tcd B及其他肿瘤标志物的检测方法应用于临床。皇冠版权所有（C）2013由爱思唯尔B. V.出版保留所有权利。
Clostridium difficile toxin B (Tcd B), as one of the primary contributing factors to the pathogenesis of C difficile-associated diseases, has raised serious public concerns due to its virulence, spore-forming ability and persistence with major types of infectious diarrhea diseases, and been used as a potential biomarker in clinical diagnoses. Thus, a simple method for the determination of Tcd B was developed based on a sandwich-type electrochemical immunosensor. Greatly enhanced sensitivity was achieved based on fabricating the immunosensor by layer-by-layer coating carbon nanotubes (MWCNTs), Prussian blue (PB), Chitosan (CS), Glutaraldehyde (GA) composite on the working electrode as well as using graphene oxide (GO) as a nanocarrier in a multienzyme amplification strategy. In comparison with conventional methods, the proposed immunoassay exhibited high sensitivity and selectivity for the detection of Tcd B, providing a better linear response range from 0.003 to 320 ng/mL and a lower limit of detection CLOD) of 0.7 pg/mL (S/N=3) under optimal experimental conditions. The immunosensor exhibited convenience, low cost, rapidity, good specificity, acceptable stability and reproducibility. Moreover, satisfactory results were obtained for the determination of Tcd B in real human stool samples, indicating that the developed immunoassay has the potential to find application in clinical detection of Tcd B and other tumor markers as an alternative approach. Crown Copyright (C) 2013 Published by Elsevier B.V. All rights reserved.