双重实时荧光PCR法测定食品中大豆、小麦过敏原  

Determination of soybean and wheat allergens in food by dual real-time fluorescence PCR

作　　者：金萍 丁洪流 张敏 金晓红 JIN Ping;DING Hong-liu;ZHANG Min;JIN Xiao-hong(Suzhou Products Quality Supervision and Inspection Institute,Suzhou,Jiangsu 215104,China)

机构地区：[1]苏州市产品质量监督检验院,江苏苏州215104

出　　处：《食品与机械》2022年第9期59-63,102,共6页Food and Machinery

基　　金：苏州市科技局民生科技项目(编号:SS202038);江苏省市场监督管理局科技计划项目(编号:KJ21125075)。

摘　　要：目的:为食品监管部门有效检测食品中大豆、小麦过敏原提供技术支撑。方法:分别依据小麦醇溶蛋白基因及大豆Lectin基因为模板设计并创建TaqMan探针双重荧光PCR方法,大豆Lectin基因检测采用FAM标记,小麦醇溶蛋白基因检测采用HEX标记,同时以真核生物18S rRNA作为内参基因确保检测体系的有效性。结果:所创建的实时多重TaqMan探针PCR体系对大豆、小麦过敏原之外的物种成分无荧光扩增;大豆、小麦混合样品的检出限均能达到0.01%(质量分数)。结论:所创建的实时多重TaqMan探针PCR体系针对大豆、小麦过敏原有高特异性,可用于食品中过敏原大豆、小麦成分的同步快速检测。Objective: A multiplex fluorescence PCR method based on TaqMan probe was developed for simultaneous detection of soybean and wheat allergens in food. Methods: TaqMan probe dual PCR was designed and established based on wheat gliadin gene and soybean lectin gene, respectively. The soybean Lectin gene was detected with FAM marker, and the wheat gliadin gene was detected with HEX marker. Moreover, the eukaryotic 18 S rRNA was used as internal reference gene to ensure the effectiveness of the detection system. Results: The multiplex fluorescence PCR method for detection of soybean and wheat allergens created in this study had no cross-reaction to ingredients other organisms showing strong specificity. The detection limit of soybean and wheat mixture was up to 0.01%(mass ratio), and the detection sensitivity was high. Conclusion: The established TaqMan probe real-time multiplex detection system for soybean and wheat allergens has the characteristics of good specificity and high sensitivity, which can be used for simultaneous and rapid detection of soybean and wheat allergens in food.

关 键 词：实时荧光聚合酶链反应 食品过敏原 大豆 小麦 

分 类 号：TS207.3[轻工技术与工程—食品科学]