College of Food Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
Biosens Bioelectron. 2012 Apr 15;34(1):44-50. doi: 10.1016/j.bios.2011.12.057. Epub 2012 Jan 6.
Mycotoxins are highly toxic contaminants in food, animal feed, and commodities. The study has developed an immunochip for quantifying the concentrations of six mycotoxins: aflatoxin B1, aflatoxin M1, deoxynivalenol, ochratoxin A, T-2 toxin, and zearalenone, which were added to drinking water. The complete antigens (Ags) of the mycotoxins were contact printed and immobilized onto agarose-modified glass slides with 12 physically isolated subarrays, based on the reaction of both diffusion and covalent bond. The optimal concentration of each antigen and antibody (Ab) was obtained using an Ag-Ab immunoassay. Based on the indirect competitive immunoassay for the simultaneous detection of six mycotoxins in one single chip, six standard curves with good logistic correlation (R(2)>0.97) were respectively plotted. The working ranges (0.04-1.69, 0.45-3.90, 20.20-69.23, 35.68-363.18, 0.11-1.81, and 0.08-7.47 ng/mL, respectively) were calculated, as well as the median inhibitory concentrations (0.31±0.04, 1.49±0.21, 34.54±1.30, 134.06±11.75, 0.49±0.05, and 1.54±0.22 ng/mL, respectively), when six mycotoxins were detected simultaneously. Finally, the recovery rates in drinking water generally ranged from 80% to 120% on the same chip, with an intra-assay coefficient of variation lower than 15%. We successfully established an immunochip for simultaneous detection of six mycotoxins within 4h, with advantages of using minimal samples and being visually semiquantitative with our naked eyes. In summary, the method could be developed on one single chip for detecting multiple contaminants in actual samples.
真菌毒素是食品、动物饲料和商品中高度有毒的污染物。本研究开发了一种免疫芯片，用于定量测定六种真菌毒素的浓度：黄曲霉毒素 B1、黄曲霉毒素 M1、脱氧雪腐镰刀菌烯醇、赭曲霉毒素 A、T-2 毒素和玉米赤霉烯酮，这些毒素被添加到饮用水中。真菌毒素的完全抗原（Ag）通过扩散和共价键的反应接触印刷并固定在琼脂糖修饰的玻璃载玻片上的 12 个物理分离的亚阵列上。使用 Ag-Ab 免疫分析获得每种抗原和抗体（Ab）的最佳浓度。基于同时在单个芯片上检测六种真菌毒素的间接竞争免疫分析，分别绘制了具有良好逻辑相关（R（2）>0.97）的六个标准曲线。计算了工作范围（分别为 0.04-1.69、0.45-3.90、20.20-69.23、35.68-363.18、0.11-1.81 和 0.08-7.47 ng/mL），以及中值抑制浓度（分别为 0.31±0.04、1.49±0.21、34.54±1.30、134.06±11.75、0.49±0.05 和 1.54±0.22 ng/mL），当同时检测六种真菌毒素时。最后，在同一芯片上，饮用水中的回收率通常在 80%至 120%之间，内部分析变异系数低于 15%。我们成功地建立了一种在 4 小时内同时检测六种真菌毒素的免疫芯片，该方法具有使用最小样本和用肉眼进行半定量的优点。总之，该方法可以在单个芯片上发展，用于检测实际样品中的多种污染物。
