杨华,徐霞红,郭玉娜,肖英平,唐标,王强.基于MOCPs-MWCNTs的大肠杆菌电化学免疫传感器[J].农业机械学报,2017,48(6):328-333.
YANG Hua,XU Xiahong,GUO Yuna,XIAO Yingping,TANG Biao,WANG Qiang.Electrochemical Immunosensor Assay (EIA) of E. coli O157:H7 Based on MOCPs-MWCNTs with Highly Efficient Antibody Immobilization[J].Transactions of the Chinese Society for Agricultural Machinery,2017,48(6):328-333.
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基于MOCPs-MWCNTs的大肠杆菌电化学免疫传感器   [下载全文]
Electrochemical Immunosensor Assay (EIA) of E. coli O157:H7 Based on MOCPs-MWCNTs with Highly Efficient Antibody Immobilization   [Download Pdf][in English]
投稿时间:2017-04-20  
DOI:10.6041/j.issn.1000-1298.2017.06.043
中文关键词:  大肠杆菌O157:H7  金属有机配位聚合物  多壁碳纳米管  电化学免疫传感器
基金项目:浙江省自然科学基金项目(LZ15C170001)
作者单位
杨华 浙江省农业科学院农产品质量标准研究所 
徐霞红 浙江省农业科学院农产品质量标准研究所 
郭玉娜 浙江省农业科学院农产品质量标准研究所 
肖英平 浙江省农业科学院农产品质量标准研究所 
唐标 浙江省农业科学院农产品质量标准研究所 
王强 浙江省农业科学院农产品质量标准研究所 
中文摘要:食源致病菌引起的食品安全问题已成为全世界关注的焦点。设计了一种基于多壁碳纳米管(MWCNTs)-有机配位聚合物(MOCPs)高效固定抗体的大肠杆菌电化学免疫传感器。利用NaAuCl4与1,4-苯二硫醇(BDT)反应生成金属有机配位聚合物,同时在BDT的还原作用下生成大量纳米金,并包埋大量吸附在MWCNTs表面的大肠杆菌抗体,最终将抗体高效包埋在MOCPs-MWCNTs中,基于MOCPs-MWCNTs固定抗体研制修饰电极,在目标物大肠杆菌O157:H7的存在下,辣根过氧化氢酶标记的抗体(HRP-Ab)随后特异性捕获于电极表面形成典型夹心结构,通过HRP催化底物产生电化学信号从而实现大肠杆菌O157:H7的检测。MOCPs-MWCNTs特有的高比表面积高效固定了大量抗体,增加了传感器结合抗体的效率并提高了传感器的灵敏度。该传感器对大肠杆菌O157:H7的检测线性范围为67~6.7×106cfu/mL,最低检出限为40cfu/mL。
YANG Hua  XU Xiahong  GUO Yuna  XIAO Yingping  TANG Biao  WANG Qiang
Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences,Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences,Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences,Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences,Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences and Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences
Key Words:Escherichia coli O157:H7  metal organic coordination polymers  multi-walled carbon nanotubes  electrochemical immunosensor
Abstract:Microbiological contamination caused by food borne diseases has become a major public health problem of the world. A novel electrochemical immunosensor assay (EIA) for high sensitive and specific detection of Escherichia coli O157:H7 was developed. A new nanocomposites with multi walled carbon nanotubes (MWCNTs) embedded inmetal organic coordination polymers (MOCPs) was successfully prepared as highly efficient matrices of capturing antibody immobilization for sensitive electrochemical biosensing. In the presence of target E. coli O157:H7, horse radish peroxidase (HRP)labeled antibody was captured on the electrode surface to form a sandwich type system via the specific identification. As a result, E. coli O157:H7 detection was realized by outputting a redox current from electro reduction of hydrogen peroxide reaction catalyzed by HRP. In the assay, the combination of the unique properties of MWCNTs and MOCPs can not only accelerate electron transfer on the electrode interface, but also provide an excellent scaffold for the conjugation of capture antibody. Meanwhile, adopting the MWCNTs-MOCPs materials significantly improved the target capturing efficiency and enhanced the sensitivity of the biosensor. The results revealed that the calibration plot obtained for E. coli O157:H7 was approximately linear from 67 cfu/mL to 6.7×106 cfu/mL with the limit of detection of 40cfu/mL. In addition, the biosensor was successfully applied to quantitative assay of E. coli O157:H7 in synthetic sample (milk). Hence, the developed electrochemical based immunosensor might provide a useful and practical tool for E. coli O157:H7 determination and related food safety analysis and clinical diagnosis.

Transactions of the Chinese Society for Agriculture Machinery (CSAM), in charged of China Association for Science and Technology (CAST), sponsored by CSAM and Chinese Academy of Agricultural Mechanization Science(CAAMS), started publication in 1957. It is the earliest interdisciplinary journal in Chinese which combines agricultural and engineering. It always closely grasps the development direction of agriculture engineering disciplines and the published papers represent the highest academic level of agriculture engineering in China. Currently, nearly 8,000 papers have been already published. There are around 3,000 papers contributed to the journal each year, but only around 600 of them will be accepted. Transactions of CSAM focuses on a wide range of agricultural machinery, irrigation, electronics, robotics, agro-products engineering, biological energy, agricultural structures and environment and more. Subjects in Transactions of the CSAM have been embodied by many internationally well-known index systems, such as: EI Compendex, CA, CSA, etc.

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