可视化膜芯片技术检测常见食源性过敏原成分

目的 建立包括鱼、虾、鸡、鸭、花生、核桃、小麦以及大豆过敏原成分的基因膜芯片技术, 实现对这几种食源性过敏原的同步可视化检测。方法 针对常见食源性过敏物质的成分特异性基因或者过敏原蛋白基因设计特异性的带生物素标记的引物和探针。采用反向斑点杂交(reversedot blot, RDB)结合多重聚合酶链式反应(multiplex polymerase chain reaction, MPCR)技术, 最终通过化学显色直观显示检测结果, 实现对多种食源性过敏原的同步可视化检测。结果 所建立的可视化基因膜芯片方法准确性强, 通过单目标物以及多目标物特异性实验, 显示本方法仅对鱼、虾、花生、核桃、大豆等靶标食源性过敏原有特异性反应, 非靶标物检测均为阴性; 通过制备不同质量浓度比的模拟样品考核方法的灵敏度, 经检测, 方法检测灵敏度可达0.1%(质量分数)。结论 所建立的方法可达到可视化、快速、准确地鉴别食品中鱼、虾、花生、核桃、大豆等常见食源性过敏原。

Detection of common food-borne allergens components by visual membrane chip technology

Objective To establish a gene membrane microarray chip technology including fish, shrimp, chicken, duck, peanut, walnut, wheat and soybean allergens, and realize the simultaneous visual detection of these food-borne allergens. Methods Specific biotin-labeled primers and probes were designed for the component specific genes or allergen protein genes of common food-borne allergens. Reversedot blot (RDB) technique combined with multiplex polymerase chain reaction (MPCR) technique was used. Finally, the detection results were visually displayed by chemical color development to realize the synchronous visual detection of multiple food-borne allergens. Results The established visualization gene microarray chip technology had high accuracy. Through the single target and multi-target specificity experiments, it was shown that this method was only specific for fish, shrimp, peanut, walnut, soybean and other target food-borne allergens, and the detection of non-target was negative. The sensitivity of the method was tested by preparing simulated samples with different mass concentration ratios. The sensitivity of the method was up to 0.1% (mass fraction). Conclusion The established method can achieve visual, rapid and accurate identification of food-borne allergens in food, such as fish, shrimp, peanut, walnut, soybean, etc..