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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。
Objective To establish a real-time fluorescent quantitative PCR assay for the allergen composition of hazelnut, and compare the method with enzyme-linked immunosorbent assay (ELISA). Methods According to the oleosin specific gene of hazelnut, we designed and screened the appropriate primers and probes, optimized the reaction system and reaction conditions, and established the real-time fluorescent quantitative PCR detection method of hazelnut allergen. The results of fluorescence quantitative PCR and ELISA analysis. Results The hazelnut allergen composition established by real-time fluorescence quantitative PCR was of good specificity and could be used for the quantitative detection of hazelnut allergen. However, the detection sensitivity was lower than that of ELISA. Conclusion The established Hazelnut allergen composition of fluorescence quantitative PCR detection method has good specificity and sensitivity of 10 mg / kg, which has good practicability. The sensitivity of ELISA detection method is higher than that of fluorescence quantitative PCR method, but when hazelnut allergy protein False negative results may occur after being destroyed.