Evaluation and Optimization of Three Different Immunoassays for Rapid Detection Zearalenone in Fodders
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Evaluation and Optimization of Three Different Immunoassays for Rapid Detection Zearalenone in Fodders Xuemei Jiang 1,2 & Xiumei Li 2 & Zhi Yang 2 & Sergei A. Eremin 3 & Xiaoying Zhang 1
Received: 19 February 2016 / Accepted: 12 June 2016 # Springer Science+Business Media New York 2016
Abstract Zearalenone (ZEN) is one of the most common contaminants in fodder with obvious reproduction toxicity and potential carcinogenicity to animals and humans. Thus, simple and sensitive methods are required for the detection of ZEN. In this work, the anti-ZEN monoclonal antibody (mAb) was prepared by hybridoma technique. Then, the mAb-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), fluorescence polarization immunoassay (FPIA), and immunochromatographic assay (ICA) were evaluated and optimized for ZEN detection in spiked fodder samples. The ic-ELISA and FPIA showed recovery ranges from 80 to 100 %. The limit of detection (LOD) in ic-ELISA, FPIA and ICA were 0.06, 0.54, and 10 ng/mL, respectively. The detection ranges of IC20~IC80 were 2.89 to 115.36 ng/mL in ic-ELISA and 3.0 to 1052 ng/mL in FPIA. Amongst three immunoassays, the ic-ELISA was the most sensitive and stable, nevertheless, most time-consuming with narrower detection range. The FPIA showed good sensitivity, precision, and wide detection range, but it required specific tracer. ICA was a time-saving handy method but exhibited relatively lower precision and quantification compared to other two methods. Keywords Zearalenone (ZEN) . Monoclonal antibody (mAb) . Indirect competitive enzyme-linked immunosorbent
* Xiaoying Zhang [email protected] 1
College of Veterinary Medicine, Northwest A&F University, Post Box 19, Xinong Rd. 22, Yangling, Shaanxi Province 712100, China
2
Laipson Health Information Technology Co., Ltd, He’nan, Luoyang 471000, China
3
Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov State University, 119992 Moscow, Russia
assay (ic-ELISA) . Fluorescence polarization immunoassay (FPIA) . Immunochromatographic assay (ICA)
Introduction Zearalenone (ZEN), as one of the commonest contaminants in fodder, has been reported to be reproduction toxic and potential carcinogenic to animals and humans (Streit et al. 2012; Zhu et al. 2012).ZEN is the secondary metabolite produced by some Fusarium genera, which mainly contaminates grains including corn, barley, wheat, oats, and sorghum worldwide (Berthiller et al. 2013). ZEN would be produced in food grains, if it is not handled and dried properly (Chun et al. 2009). The ZEN is stable at high temperatures and could not be degraded during food processing (Mirocha et al. 2013). In order to decrease the ZEN-induced incidences, the USA and Europe have defined the permissible limit of ZEN in food products ranged from 0.02 to 1000 μg/kg (EC/No. 1881/2006). In China, the legal limit is 500 μg/ kg in formulated feed and 60 μg/kg in cereals (Zhang et al. 2013). With the focus to prevent the hazards of ZEN contaminated fodders before market entry, various methods have be
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