深加工食品中苦瓜成分实时荧光PCR定性检测方法的建立与应用

Establishment and Application of Qualitative Real-Time PCR Detection of Bitter Melon Ingredients in Highly Processed Foods

摘要: 为实现对深加工食品中苦瓜成分的真伪和品质鉴别，文章建立了深加工食品中苦瓜成分实时荧光聚合酶链式反应(polymerase chain reaction, PCR)定性检测方法。根据苦瓜基因组DNA中的保守基因，分别设计3组实时荧光PCR定性检测的特异性引物探针，筛选最优的引物探针，优化PCR反应体系，并对方法的检测低限、稳定性和市售深加工食品的适用性进行了验证。该方法特异性和稳定性良好，对苦瓜基因组DNA检测低限为0.05%，利用该方法对29批次市售深加工食品进行苦瓜成分鉴别，发现其中有1批次未检出苦瓜成分。该方法可有效鉴别各种市售深加工食品中的苦瓜成分。

Abstract: In order to achieve the authenticity and quality identification of bitter melon ingredients in highly processed food, a qualitative real-time fluorescent polymerase chain reaction (PCR) method was established. 3 sets of specific primers and probes were designed and selected according to the conserved sequences in genomic DNA, and the reaction system was optimized for the best performance. Afterwards, the limit of detection and the stability of this method, as well as its applicability in market-sale highly processed food, were verified. The specificity and stability were good and the method LOD was indicated as 0.05%. The method was used to identify the ingredients of bitter melon in 29 batches of market-sale highly processed food and 1 batch was not detected to contain bitter gourd components. The method is effective for the identification of bitter melon ingredients in market-sale highly processed food.