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用于大肠杆菌(作为与乳制品质量相关的指示菌)的放大电化学免疫分析的功能化金纳米棒标记物。

Functionalized gold nanorod-based labels for amplified electrochemical immunoassay of E. coli as indicator bacteria relevant to the quality of dairy product.

作者信息

Zhang Xinai, Zhang Fan, Zhang Hongyin, Shen Jianzhong, Han En, Dong Xiaoya

机构信息

School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China.

Liaocheng People׳s Hospital, Liaocheng 252002, PR China.

出版信息

Talanta. 2015 Jan;132:600-5. doi: 10.1016/j.talanta.2014.10.013. Epub 2014 Oct 22.

Abstract

In this paper, we report an amplified electrochemical immunoassay for Escherichia coli as indicator bacteria relevant to the quality of dairy product using the functionalized gold nanorod-based labels ({dAb-AuNR-FCA}). The {dAb-AuNR-FCA} labels were designed by exploiting silica-functionalized gold nanorods (AuNR@SiO2) as the carriers for immobilization of detection antibody (dAb) and ferrocenecarboxylic acid (FCA), in which dAb was used for recognition of E. coli and FCA tags served as signal-generating molecule. Greatly amplified signal was achieved in the sandwich-type immunoassay when enormous FCA linked to AuNR@SiO2. Compared with the commercially available {dAb-FCA}, the {dAb-AuNR-FCA} labels exhibited a better performance for E. coli assay due to the advantages of AuNR@SiO2 as carriers. Under optimal experimental conditions, it showed a linear relationship between the peak current of FCA and the logarithmic value of E. coli concentration ranging from 1.0×10(2) to 5.0×10(4) cfu mL(-1) with a detection limit of 60 cfu mL(-1) (S/N=3), and the electrochemical detection of E. coli could be achieved in 3h. Moreover, the proposed strategy was used to determine E. coli in dairy product (pure fresh milk, yogurt in shelf-life, and expired yogurt), and the recoveries of standard additions were in the range of 95.1-106%. This proposed strategy exhibited rapid response, high sensitivity and specificity for E. coli assay in dairy product, and could become a promising technique to estimate the quality of dairy product.

摘要

在本文中,我们报道了一种使用功能化金纳米棒基标记物({dAb-AuNR-FCA})对与乳制品质量相关的指示菌大肠杆菌进行放大电化学免疫分析的方法。{dAb-AuNR-FCA}标记物是通过利用二氧化硅功能化金纳米棒(AuNR@SiO2)作为载体来固定检测抗体(dAb)和二茂铁羧酸(FCA)设计而成的,其中dAb用于识别大肠杆菌,FCA标签作为信号产生分子。当大量FCA与AuNR@SiO2相连时,在夹心型免疫分析中实现了信号的大幅放大。与市售的{dAb-FCA}相比,{dAb-AuNR-FCA}标记物由于AuNR@SiO2作为载体的优势,在大肠杆菌检测中表现出更好的性能。在最佳实验条件下,FCA的峰值电流与大肠杆菌浓度的对数在1.0×10(2)至5.0×10(4) cfu mL(-1)范围内呈线性关系,检测限为60 cfu mL(-1)(S/N=3),并且大肠杆菌的电化学检测可在3小时内完成。此外,所提出的策略用于测定乳制品(纯鲜牛奶、保质期内的酸奶和过期酸奶)中的大肠杆菌,加标回收率在95.1-106%范围内。该所提出的策略对乳制品中大肠杆菌的检测表现出快速响应、高灵敏度和特异性,并且可能成为评估乳制品质量的一种有前景的技术。

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