基于荧光定量PCR鉴定冷鲜肉制品中羊源性成分及其含量  

Identification of Sheep-derived Components and Their Content in Cold Fresh Meat Products Based on Fluorescent Quantitative PCR

作　　者：史莹莹 邵俊锋 郭娟 许慧卿[1] SHI Ying-ying;SHAO Jun-feng;GUO Juan;XU Hui-qing(College of Food Science and Technology,Yangzhou University,Yangzhou 225127,China)

机构地区：[1]扬州大学食品科学与工程学院,江苏扬州225127

出　　处：《食品工业科技》2020年第11期146-150,共5页Science and Technology of Food Industry

摘　　要：建立一种快速、准确鉴定羊肉制品中羊源性成分并且量化羊肉成分含量的方法。以羊线粒体细胞色素b基因为靶基因,设计出具有特异性引物。选择真核生物核糖体16S rDNA为内参基因,采用实时荧光相对定量法。羊肉质量百分比的对数值与之对应的循环阈值差值ΔCt呈良好线性关系。标准曲线回归公式为y=-3.4057x-0.3112,R^2=0.9916,扩增效率达E达96.62%。本试验中羊源性DNA检测限可达16 pg/μL,回收率在93.07%~106.20%。抽取8份市售羊肉制品进行检测,有3份含量低于50%。本试验中建立的实时荧光PCR检测方法操作方便、特异性强、灵敏度高,所得数据可靠,为肉制品羊源性成分检测提供了参考方法。In order to establish a method for quickly and accurately identifying sheep-derived components in sheep products and quantifying the content of sheep components,specific primers were designed with sheep mitochondrial cytochrome b gene as the target gene.The eukaryotic ribosomal 16 S rDNA was selected as the internal reference gene,and the real-time fluorescence relative quantification method was used.The logarithmic value of the percentage of mutton mass had a good linear relationship with the corresponding cycle threshold differenceΔCt.The standard curve regression formula was y=-3.4057 x-0.3112,R^2=0.9916,and the amplification efficiency reached 96.62%.In this test,the detection limit of sheep-derived DNA could reach 16 pg/μL,and the recovery rate was between 93.07%and 106.20%.Taking 8 commercial mutton products for testing,3 of them were below 50%.The real-time fluorescence PCR detection method established in this experiment was convenient to operate with strong specificity,high sensitivity,and reliable data,which provided a reference method for the detection of sheep-derived components in meat products.

关 键 词：实时荧光定量PCR 羊肉 线粒体细胞色素B 相对定量 

分 类 号：TS201.3[轻工技术与工程—食品科学]