实时荧光PCR技术快速检测奶制品中掺入的大米源性成分  

Detection for rice-derived ingredients present in dairy products with real-time PCR method

作　　者：刘小艳[1] 傅春玲[1] 李培[2] 丁洪流[2] 金萍[2] 

机构地区：[1]苏州大学医学部公共卫生学院,江苏苏州215123 [2]苏州市产品质量监督检验所,江苏苏州215128

出　　处：《食品工业科技》2014年第3期302-304,335,共4页Science and Technology of Food Industry

基　　金：苏州市科技支撑计划项目(SS201126)

摘　　要：目的:建立基于实时荧光PCR技术的奶制品中掺入大米源性成分的快速检测方法。方法:以水稻的根部表达基因(gos9)为靶基因设计特异性引物和探针,经特异性实验和灵敏度实验验证引物探针可行性,并经模拟含大米奶粉及市售奶类制品检测验证其实际检测能力。结果:该引物探针体系只针对大米DNA进行扩增,与奶类主成分牛、羊及其它谷类和植物性食物DNA均无交叉扩增;最低能检测到0.1ng的大米DNA,对含大米粉的模拟混合奶粉样品,检出限可达0.1%(W/W)。将其应用于21份市售奶制品样品检测,对含大米源性成分的奶制品扩增阳性,检测结果与食品标签相符。结论:该实时荧光PCR检测体系具有快速、特异、灵敏的优点,可以准确鉴定出奶制品中大米成分,适用于奶类中掺加大米源性成分的检测。Objective：This study aimed to establish a real-time PCR method for detection of rice-derived ingredients present in dairy products, Method= Primers and Taqman probe of this assay were designed for specific amplification of rice DNA targeting its root-specific gene gos9 gene.Specificity and sensitivity test was performed to evaluate its utility.The assay was further validated with mixtures of milk powder containing various rice powder and commercial dairy products samples with and without rice ingredients from retail market.Results：The assay was proved to be highly specific for rice with no cross-reaction to bovine, ovine and close-related cereal and plant food ingredients may occur in dairy products.The detection limit was 0.1 ng for pure rice DNA and 0.t % （W/W） for rice flour spiked in whole milk powder, respectively. When applied to detect 21 commercial samples, it yields positive amplification signals for the samples with rice ingredients while negative for those without ones demonstrating corresponded results identical to the species indicated in the food label. Conclusions：The assay provided a fast and reliable mean of potential advantages in routine determination of dairy products for the presence of rice-derived ingredients.

关 键 词：实时荧光PCR 大米 gos9基因 奶制品 

分 类 号：TS207.3[轻工技术与工程—食品科学]