Quantitative Determination of Deoxynivalenol (DON) Using the Amplified Luminescent Proximity Homogeneous Assay (AlphaLIS
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Quantitative Determination of Deoxynivalenol (DON) Using the Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA) Zhou Bin & Wang Ke & Jin Jian & Chen Yun & Huang Biao & Zhao Weiguo
Received: 6 January 2010 / Accepted: 27 April 2010 / Published online: 12 May 2010 # Springer Science+Business Media, LLC 2010
Abstract An indirect competitive Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA) was established by using anti-deoxynivalenol (DON) polyclonal antibody and coating antigen DON-bovine serum albumin (BSA) for detection of DON in cereals. DON-BSA was coated on Acceptor bead, the kit also contains donor bead sensitizer particles coated with streptomycin. The optional test conditions and analytical performance of the method were studied. Working concentration ranged from 0.007 to 100 ng/ml and the sensitivity for detection was 0.007 ng/ml. The intra- and inter-batch coefficient of variation (CV) of the assay were both below 5%. The recovery rate of artificially contaminated cereals ranged from 81.1% to 110.9% while the mean recovery of DON from cereals was 94.4%. This study suggests that DON-AlphaLISA method is a good method with high sensitivity and rapidity for quantitative analyzing DON in cereals. Keywords Deoxynivalenol (DON) . Biotoxin . Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA)
Z. Bin : W. Ke : H. Biao (*) Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China e-mail: [email protected] J. Jian : C. Yun School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, Jiangsu 214063, China Z. Weiguo Shanghai Boyang Biotechnology Company Ltd, Shanghai 210000, China
Introduction Deoxynivalenol (DON, also called vomitoxin) is a trichothecene mycotoxin produced by various species of fungi. It is a mycotoxin commonly found in corn, wheat, bread, barley, as well as other most important sources of human intake (Wu 2004). Contamination of foods by DON is a potential hazard for livestock and human beings. DON’s emetic and anorectic potencies are greater than those reported for the more acutely toxic trichothecenes. Toxicological effects associated with DON poisoning in humans and livestock include reducing weight gain, emesis, and gastroenteritis (Zhang et al. 1998; Vesely and Vesela 1995). The immune system is extremely sensitive to trichothecenes. Exposure to low DON doses induces rapid transient up-regulation of proinflammatory cytokines causing immune stimulation, whereas high doses of DON cause apoptosis in lymphoid tissues resulting in immunosuppression (Chuang et al. 2007). It is therefore of utmost importance to assay DON contamination. Various analytical methods have been developed for the detection of DON contamination in cereals. Immunoassays as rapid, low cost, and high throughput tests are convenient for rapid screening of a large number of samples. Enzyme-linked immunosorbent assays (ELISA) have been widely reported for the analysis of D
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