Zhang Xiya, Wu Chao, Wen Kai, Jiang Haiyang, Shen Jianzhong, Zhang Suxia, Wang Zhanhui
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Molecules. 2015 Dec 28;21(1):E27. doi: 10.3390/molecules21010027.
A new highly specific and sensitive monoclonal antibody (MAb) to T-2 toxin (T-2) was produced, providing an IC50 value of 1.02 ng/mL and negligible cross-reactivity (CR) to other related mycotoxins. Based on the new MAb, a lateral-flow immunochromatographic assay (LFIA) using colloidal gold (CG) and fluorescent microspheres (FMs) as labels was proposed for T-2. Under the optimized conditions, in rapid qualitative assay, the cut-off values of the CG-LFIA were 400 μg/kg in rice and 50 μg/L in fresh milk, and the cut-off values of the FMs-LFIA were 100 μg/kg in both rice and chicken feed. For the quantitative assay with the FMs-LFIA, the limit of detection (LOD) were 0.23 μg/kg and 0.41 μg/kg in rice and chicken feed, respectively, and the average recoveries ranged from 80.2% to 100.8% with the coefficient of variation (CV) below 10.8%. In addition, we found that the CG-LFIA could tolerate the matrix effect of fresh milk better than the FMs-LFIA, while the FMs-LFIA could tolerate the matrix effect of chicken feed better than CG-LFIA under the same experimental conditions. These results provide a certain reference for the selection of appropriate labels to establish a rapid LFIA in various biological samples.
制备了一种针对T-2毒素（T-2）的新型高特异性和高灵敏度单克隆抗体（MAb），其IC50值为1.02 ng/mL，与其他相关霉菌毒素的交叉反应（CR）可忽略不计。基于这种新型单克隆抗体，提出了一种以胶体金（CG）和荧光微球（FMs）为标记物的用于检测T-2的侧向流动免疫色谱分析（LFIA）方法。在优化条件下，快速定性分析中，CG-LFIA在大米中的截断值为400 μg/kg，在鲜牛奶中的截断值为50 μg/L，FMs-LFIA在大米和鸡饲料中的截断值均为100 μg/kg。对于FMs-LFIA的定量分析，大米和鸡饲料中的检测限（LOD）分别为0.23 μg/kg和0.41 μg/kg，平均回收率在80.2%至100.8%之间，变异系数（CV）低于10.8%。此外，我们发现，在相同实验条件下，CG-LFIA比FMs-LFIA更能耐受鲜牛奶的基质效应，而FMs-LFIA比CG-LFIA更能耐受鸡饲料的基质效应。这些结果为选择合适的标记物以在各种生物样品中建立快速LFIA提供了一定参考。
