荧光定量PCR方法检测榛果过敏原成分

Detection of the allergen of hazelnut by real-time PCR

中文关键词:  榛果  过敏原  荧光定量PCR  酶联免疫吸附法

英文关键词:hazelnut  allergen  real-time PCR  enzyme-linked immunosorbent assay

中文摘要:

      目的 建立榛果过敏原成分的荧光定量PCR检测方法, 并将此方法与酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)方法进行比对实验。方法 根据榛果成分oleosin特异性基因设计并筛选合适的引物和探针, 优化反应体系和反应条件, 建立榛果过敏原成分的荧光定量PCR检测方法, 对荧光定量PCR方法与ELISA方法检测结果进行分析。结果 建立的榛果过敏原成分荧光定量PCR方法特异性良好, 可用于榛果过敏原成分的定量检测, 但检测灵敏度低于ELISA检测方法。结论 所建立的榛果过敏原成分的荧光定量PCR检测方法特异性好, 灵敏度达到10 mg/kg, 具有较好的实用性, ELISA检测方法灵敏度高于荧光定量PCR法, 但当榛果过敏蛋白被破坏后有可能出现假阴性结果。

英文摘要:

      Objective To establish a real-time PCR method for detection of the allergen of hazelnut, and compare the method with the enzyme-linked immunosorbent assay (ELISA) method. Methods According to the oleosin gene of hazelnut, the optimized reaction system and the reaction conditions had been proposed. To establish a real-time PCR method for detection of the allergen of hazelnut, a comparative analysis of the detection results by this method and ELISA was made. Results The method was specific for hazelnut allergens, and was suitable for the detection of hazelnut allergens. But the sensitivity of this method was lower than ELISA. Conclusion The real-time PCR method for detecting the allergen of hazelnut is suitable for application, the sensitivity is up to 10 mg/kg, lower than the ELISA detection method, but a false negative result may occur in ELISA method due to the hazelnut allergic protein damaged.