A disposable amperometric enzyme immunosensor for rapid detection of Vibrio parahaemolyticus in food based on agarose/Nano-Au membrane and screen-printed electrode
A disposable amperometric enzyme immunosensor for rapid detection of Vibrio parahaemolyticus in food based on agarose/Nano-Au membrane and screen-printed electrode
复制标题
DOI：
10.1016/j.elecom.2007.01.036
复制
发表时间：
2007-06
影响因子：
5.4
通讯作者：
Guangying Zhao;Fengfeng Xing;Shaoping Deng
中科院分区：
文献类型：
--
作者：
Guangying Zhao;Fengfeng Xing;Shaoping Deng
最近爆发的食源性疾病继续支持需要快速和灵敏的方法来检测食源性病原体。本文研制了一种检测副溶血性弧菌(VP)的一次性电化学传感器，该传感器以琼脂糖凝胶/纳米金膜为电极，辣根过氧化物酶(HRP)标记的副溶血性弧菌抗体(HRP-anti-VP)为传感器。用循环伏安法(CV)和激光扫描共聚焦显微镜(LSCM)对该免疫传感器进行了表征。免疫传感器在室温(25±0.5℃)下与VP一步法孵育30min。VP通过免疫反应连接到免疫传感器表面，部分抑制了HRP活性中心对H_2O_2氧化硫氨酸反应的进入。根据还原电流的漂移可以定量检测VP，而循环伏安法作为检测酶反应产物的电化学手段。在最佳的免疫反应和电化学检测条件下，用S型曲线法快速检测VP，线性范围为105~109cfu/ml，检出限为7.374×104cfu/ml(S/N=3)。该免疫传感器具有良好的特异性、重现性、稳定性和准确性，能够满足实际样品检测的需要。
Recent outbreaks of foodborne illnesses continue to support the need for rapid and sensitive methods for detection of foodborne pathogens. A disposable electrochemical immunosensor for detection of Vibrio parahaemolyticus (VP) based on the screen-printed electrode (SPE) coated with agarose/Nano-Au membrane and horseradish peroxidase (HRP) labeled VP antibody (HRP-anti-VP) has been developed in this paper. Then, the immunosensor was characterized by cyclic voltammetry (CV) and laser scanning confocal microscope (LSCM). The immunosensor was incubated with the one-step immunoassay format involving VP for 30min at room temperature (25±0.5°C). The access of the active center of HRP catalyzing the oxidation reaction of thionine by H2O2was partly inhibited by VP, which connected on the surface of the immunosensor by immunoreaction. VP could be quantificationally detected according to the shift of reduction current while CV was used as electrochemical means to detect the products of the enzymatic reaction. Under the optimum conditions of immunoreaction and electrochemical detection, VP was rapidly detectable by sigmoidal curve method and form a linear calibration between 105and 109cfu/ml with an associated detection limit of 7.374×104cfu/ml (S/N=3). The immunosensor had acceptable specificity, reproducibility, stability and accuracy, indicating that the immunosensor could satisfy the need of practical sample detection.