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Toxins 2018, 10(5), 196; https://doi.org/10.3390/toxins10050196

Electrochemical Immunosensor for Detection of Aflatoxin B1 Based on Indirect Competitive ELISA

1
Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
2
Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
3
Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
4
Functional Devices Laboratory, Institute of Advanced Technology, Universiti Putra Malaysia, 43400 Serdang Selangor, Malaysia
*
Author to whom correspondence should be addressed.
Received: 28 March 2018 / Revised: 3 May 2018 / Accepted: 8 May 2018 / Published: 11 May 2018
(This article belongs to the Special Issue Advanced Sensors for Toxins)
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Abstract

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. View Full-Text
Keywords: indirect competitive ELISA; electrochemical immunosensor; aflatoxin B1; multi-walled carbon nanotubes; chitosan; screen-printed carbon electrode; peanut indirect competitive ELISA; electrochemical immunosensor; aflatoxin B1; multi-walled carbon nanotubes; chitosan; screen-printed carbon electrode; peanut
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Azri, F.A.; Sukor, R.; Selamat, J.; Abu Bakar, F.; Yusof, N.A.; Hajian, R. Electrochemical Immunosensor for Detection of Aflatoxin B1 Based on Indirect Competitive ELISA. Toxins 2018, 10, 196.

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