Peng Dapeng, Chang Fangfang, Wang Yulian, Chen Dongmei, Liu Zhenli, Zhou Xiaodong, Feng Liang, Yuan Zonghui
a National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety , Huazhong Agricultural University , Wuhan , Hubei , China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2016;33(4):683-92. doi: 10.1080/19440049.2016.1160153. Epub 2016 Mar 23.
The consumption of food or feed contaminated with high levels of T-2 toxin may cause adverse health effects in humans and other animals. In this study, to monitor T-2 toxin rapidly in food and feed, a sensitive and specific monoclonal antibody (mAb) against T-2 toxin was generated and a simple and rapid indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) developed. T-2 toxin was first converted to T-2-hemisuccinate (T-2HS) and T-2-hemiglutarate (T-2HG), which were then conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare an immunogen and coating antigen, respectively. After the inoculation of female Balb/c mice and cell fusions, one cell line, 4D8, with the IgG1 isotype was obtained. The 4D8 antibody exhibited the ability specifically to recognise T-2 toxin with IC50 1.46 µg l(-1). Based on this 4D8 mAb, an optimised ic-ELISA protocol was developed using only methanol-water (7:3, v/v) in feed and cereal samples and ethyl acetate in muscle samples. The limits of detection of T-2 toxin in various sample matrices varied from 0.07 to 15.8 µg kg(-1); the recoveries ranged from 50.3% to 113.6%; and the CVs were less than 19.0%. These results suggest that the prepared mAb and the developed ic-ELISA method will be a useful tool for detecting T-2 toxin in foods and feeds.
食用受高水平T-2毒素污染的食品或饲料可能会对人类和其他动物的健康产生不利影响。在本研究中，为了快速监测食品和饲料中的T-2毒素，制备了一种针对T-2毒素的灵敏且特异的单克隆抗体（mAb），并开发了一种简单快速的间接竞争酶联免疫吸附测定法（ic-ELISA）。T-2毒素首先被转化为T-2-半琥珀酸酯（T-2HS）和T-2-半戊二酸酯（T-2HG），然后分别与牛血清白蛋白（BSA）和卵清蛋白（OVA）偶联，以制备免疫原和包被抗原。在接种雌性Balb/c小鼠并进行细胞融合后，获得了一株IgG1亚型的细胞系4D8。4D8抗体表现出特异性识别T-2毒素的能力，IC50为1.46 μg l(-1)。基于该4D8单克隆抗体，开发了一种优化的ic-ELISA方法，饲料和谷物样品仅使用甲醇-水（7:3，v/v），肌肉样品使用乙酸乙酯。不同样品基质中T-2毒素的检测限在0.07至15.8 μg kg(-1)之间；回收率在50.3%至113.6%之间；变异系数小于19.0%。这些结果表明，所制备的单克隆抗体和开发的ic-ELISA方法将成为检测食品和饲料中T-2毒素的有用工具。
