竞争性酶联适配体可视化检测玉米赤霉烯酮

Visual Detection of Zearalenone Based on Competitive Emzyme-Linked Aptamer Assay

摘要:

建立一种基于竞争性酶联适配体检测食品中玉米赤霉烯酮的可视化分析方法。将生物素标记的玉米赤霉烯酮适配体通过生物素-亲和素连接固定在微孔板上，玉米赤霉烯酮适配体的互补链（cDNA）与辣根过氧化物酶（HRP）连接形成cDNA-HRP信号探针，cDNA-HRP信号探针和靶标竞争与适配体结合，加入TMB显色液和硫酸终止液后，即可实现玉米赤霉烯酮的可视化检测。在最佳反应条件下，方法最低检测限为0.7 ng/mL，在1～10 000 ng/mL范围内线性良好（R²=0.991 3），与类似的真菌毒素相比具有较高的特异性，对啤酒和玉米样品的加标回收率分别为88.57%～102.14%和91.43%～106.43%。该方法灵敏度高、特异性好且检测成本低，适用于食品中玉米赤霉烯酮的可视化检测。

Abstract:

A visual detection of zearalenone in food by competitive enzyme-linked aptamer assay was developed. A biotinylated zearalenone aptamer was immobilized on the surface of microplate wells by biotin-avidin binding. The zearalenone aptamer's complementary(cDNA) was ligated with horseradish peroxidase (HRP) to form a cDNA-HRP signal probe. Then the cDNA-HRP signal probe and target competitively combined with the aptamer. The visual detection of zearalenone could be achieved after the addition of TMB chromogenic reagent and stop solution(sulfuric acid). Under the optimized conditions, the detection limit was 0.7 ng/mL and the linearity was good in the range of 1 to 10 000 ng/mL(R²=0.991 3). Compared with other similar mycotoxins, the developed method had good specificity to zearalenone. And the spike recoveries of beer and corn samples were 88.57%～102.14% and 91.43%～106.43%, respectively. The method has high sensitivity, good specificity and low detection cost, which is suitable for the visual detection of zearalenone in food.