基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌  

Rapid Detection of Staphylococcus aureus by An Electrochemical Immunosensor Based on Egg Yolk Antibody-Metal Organic Framework Composite Probe

作　　者：王家琪 吴佳怡 林建原[3] 李天华[1] 李登峰[2] 干宁[1] WANG Jia-Qi;WU Jia-Yi;LIN Jian-Yuan;LI Tian-Hua;LI Deng-Feng;GAN Ning(School of Materials Science and Chemical Engineering,Ningbo University,Ningbo 315211,China;College of Oceanography,Ningbo University,Ningbo 315211,China;College of Biology and Environment,Zhejiang Wanli University,Ningbo 315200,China)

机构地区：[1]宁波大学材料科学与化学工程学院,宁波315211 [2]宁波大学海洋学院,宁波315211 [3]浙江万里学院生物与环境分院,宁波315200

出　　处：《分析化学》2021年第2期197-206,共10页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.21974074);浙江省自然科学基金项目(Nos.LY19B050001,LY18B070004,2019A610184);浙江省基础公益计划研究项目(Nos.KF2019001,LGN18H300001);宁波大学王宽诚幸福基金资助。

摘　　要：金黄色葡萄球菌(S.aureus)污染是影响食品质量安全的重要因素,因此迫切需要开发快速、简便的现场检测食品中S.aureus的分析方法。本研究采用灭活S.aureus免疫无特定病原体(SPF)蛋鸡,从产蛋中纯化出卵黄抗体(IgY),其可以高效、特异性地捕获S.aureus。将IgY固定在丝网印刷电极表面,可用于食品中S.aureus的选择性分离富集。在UiO-66金属有机框架(MOF)材料表面共价连接二茂铁(Fc)和苯硼酸(BPA),获得了MOF@Fc-BPA复合探针。此探针与电极上的IgY共同识别并捕获S.aureus,并在电极上产生Fc的氧化还原信号。在最佳反应条件下,电化学信号强度与S.aureus浓度的对数值成正比,检测范围为10~109 cfu/mL,检出限为3 cfu/mL,检测时间为20 min。采用此传感器检测了养殖虾体内的S.aureus,检测结果与标准ELISA方法的结果一致,但检测时间明显缩短,检出限比ELISA方法低两个数量级。此传感器具有良好的现场快速筛查S.aureus的能力。Staphylococcus aureus(S.aureus)contamination is an important factor affecting food quality and safety.Therefore,it is urgent to develop a quick and easy on-site analysis method for detecting S.aureus in food.In this work,inactivated S.aureus was used to immunize specific pathogen free(SPF)hens,and the yolk antibody(IgY)was purified from the eggs wich could efficiently and specifically capture S.aureus.IgY modified on the surface of the screen-printed electrode could be used for the selective separation and enrichment of S.aureus in the sample.Ferrocene(Fc)and phenylboronic acid(BPA)were covalently connected on the surface of UiO-66 nano-metal organic framework material(MOF)to obtain a MOF@Fc-BPA composite probe.The prepared probe and the IgY on the electrode jointly identified and captured S.aureus,and then generated a redox signal of Fc on the electrode.Under the optimal reaction conditions,the electrochemical signal intensity was proportional to the logarithm of the S.aureus concentration,the detection range was 10-109 cfu/mL,the detection limit was 3 cfu/mL,and the detection time was 20 min only.In addition,we used this sensor to detect S.aureus in cultured shrimp,and the results obtained were consistent with that of standard ELISA,while the detection time was greatly shortened,and the detection limit was two orders of magnitude lower than the ELISA method.The results indicated that the immunosensor was able to quickly screen S.aureus on site.

关 键 词：卵黄抗体 金黄色葡萄球菌 二茂铁/苯硼酸-金属有机框架复合探针 电化学免疫传感器 现场快速检测 

分 类 号：TS207.4[轻工技术与工程—食品科学] O657.1[轻工技术与工程—食品科学与工程] TP212[理学—分析化学]