中文摘要：霉菌毒素是由真菌自然产生的次生有毒代谢物。霉菌毒素分析对减少受污染的食物和饲料的消费至关重要。本研究基于间接竞争酶联免疫吸附试验(ELISA)，成功研发了一种超灵敏的电化学免疫传感器，用来检测黄曲霉毒素B1（AFB1）。该免疫传感器线性范围为0.0001到10 ng/mL，检测限为0.3 pg/mL。用来分析加标花生样品中的AFB１，回收率在80%至127%之间。通过RSD值(n = 5)评估该免疫传感器的精度，重现性和重复性分别为4.78% 和2.71%。
外文摘要：Mycotoxins are the secondary toxic metabolites produced naturally by fungi. Analysis of mycotoxins is essential to minimize the consumption of contaminated food and feed. In this present work, an ultrasensitive electrochemical immunosensor for the detection of aflatoxin B₁ (AFB₁) was successfully developed based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). Various parameters of ELISA, including antigen⁻antibody concentration, blocking agents, incubation time, temperature and pH of reagents, were first optimized in a 96-well microtiter plate to study the antigen⁻antibody interaction and optimize the optimum parameters of the assay. The optimized assay was transferred onto the multi-walled carbon nanotubes/chitosan/screen-printed carbon electrode (MWCNTs/CS/SPCE) by covalent attachment with the aid of 1-Ethyl-3-(3-dimetylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS). Competition occurred between aflatoxin B₁-bovine serum albumin (AFB₁⁻BSA) and free AFB₁ (in peanut sample and standard) for the binding site of a fixed amount of anti-AFB₁ antibody. Differential pulse voltammetry (DPV) analysis was used for the detection based on the reduction peak of TMB(ox). The developed immunosensor showed a linear range of 0.0001 to 10 ng/mL with detection limit of 0.3 pg/mL. AFB₁ analysis in spiked peanut samples resulted in recoveries between 80% and 127%. The precision of the developed immunosensor was evaluated by RSD values (n = 5) as 4.78% and 2.71% for reproducibility and repeatability, respectively.
外文关键词：indirect competitive ELISA；electrochemical immunosensor；aflatoxin B-1；multi-walled carbon nanotubes；chitosan；screen-printed carbon electrode；peanut
作者：Azri, FA；Sukor, R；Selamat, J；Abu Bakar, F；Yusof, NA；Hajian, R
作者单位：Univ Putra Malaysia
期刊名称：TOXINS
期刊影响因子：3.571
出版年份：2018
出版刊次：5
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