食品过敏原大豆成分LAMP实时浊度检测法的建立

Development of a real-time turbidimeter-based LAMP method for detection of allergen soybean in food

根据过敏原大豆的保守内源基因Lectin的序列,设计6条特异性LAMP引物,利用实时浊度仪对反应体系中扩增产物的实时监控筛选出最佳引物,建立食品中过敏原大豆成分的环介导等温扩增(LAMP)检测方法,并对该方法的特异性、灵敏度、稳定性进行了评价。结果表明:该方法能够特异性检出食品中的过敏原大豆成分,特异性高,灵敏度达0.1%(w/w)。对含有大豆成分为10%、1%、0.1%、0%(w/w)的样品分别进行20次重复实验,其稳定性好,假阳性和假阴性率为0%。本实验建立的LAMP方法适用于特异性快速检测食品中的过敏原大豆成分。

According to the endogenous gene of allergen soybean Lectin, six specific LAMP primers were designed. A loop-mediated isothermal amplification（LAMP） method for detecting allergen soybean in food was established. Real-time monitoring of the LAMP reaction was achieved by a turbidimeter. Specificity,sensitivity,stability and repeatability of this method were tested. The results showed that the method could specifically detect the soybean in food,and the detection sensitivity of the method was 0.1%（w/w）. With the soybean sample of 10%,1%,0.1%,0%（w/w） concentration as templates,stability and repeatability testing was conducted,false negative rate was 0%. The results showed that the LAMP method is suitable for specifically detecting allergen soybean in food.