摘要
目的建立一种基于TaqMan探针-实时荧光聚合酶链式反应(polymerase chain reaction,PCR)的方法,检测羊肉卷中是否掺入鸭源性成分,并定量分析两者的比例。方法以羊线粒体细胞色素b基因为靶基因,设计了羊特异性引物和荧光标记探针,以常见畜禽肉包括牛肉、羊肉、猪肉、鸡肉和鸭肉等阳性DNA为参考模板分别对待测样品(羊肉卷)进行TaqMan探针-实时荧光PCR扩增,又通过绘制标准曲线和确定羊和鸭的质量与DNA比值常数,对配比的混合肉样进行相对定量分析从而进一步确定待测样品含量。结果定性检测结果显示阳性样品和待测样品均呈现明显扩增曲线,但某农贸市场低价位(Y3)羊肉卷被检测出除羊肉之外的鸭肉成分;相对定量检测结果显示待测样品中羊肉相对含量为1.35 mg,鸭肉相对含量为0.24×10^(-5) mg。结论本研究建立的方法能有效检测羊肉卷中鸭源性成分及其相对含量。
Objective To establish a TaqMan probe-based real-time fluorescent polymerase chain reaction(PCR)method for detecting duck derived components in mutton rolls and quantitatively analyze their proportions.Methods Using sheep mitochondrial cytochrome b gene as the target gene,sheep-specific primers and fluorescent-labeled probes were designed.Positive DNA samples from common livestock and poultry meats,including beef,mutton,pork,chicken and duck,were used as reference templates to perform TaqMan probe-based real-time PCR amplification on the test samples(mutton rolls).Standard curves were plotted,and the mass-to-DNA ratio constants for sheep and duck were determined to conduct relative quantitative analysis of mixed meat samples,thereby further determining the content of the test samples.Results Qualitative detection showed significant amplification curves in both positive and test samples,but a low-priced mutton roll(Y3)from a farmers’market was found to contain duck-derived components in addition to mutton.Relative quantitative analysis revealed that the mutton content in the test sample was 1.35 mg,while the duck content was 0.24×10^(-5) mg.Conclusion The method established in this study can effectively detect duck derived components and their relative content in mutton rolls.
作者
谢海桃
陈曦
金净
查娜
鲁丽花
乔奇
陈翔
XIE Hai-Tao;CHEN Xi;JIN Jing;CHA Na;LU Li-Hua;QIAO Qi;CHEN Xiang(M-Grass Ecology and Environment(Group)Co.,Ltd.,Hohhot 010050,China;National Center of Pratacultural Technology Innovation(Under Preparation),Hohhot 010050,China;Inner Mongolia Autonomous Region Product Quality Inspection Institute,Hohhot 010010,China;National Dairy and Meat Product Quality Inspection Center Inner Mongolia,Hohhot 010010,China)
出处
《食品安全质量检测学报》
2025年第18期274-281,共8页
Journal of Food Safety and Quality
基金
内蒙古自治区科技重大专项(2024JBGS0011-4-4)
内蒙古自治区“英才兴蒙”工程团队项目(2025TYL13)。
关键词
动物源性成分
特异性引物
循环阈值
扩增曲线
实时荧光聚合酶链式反应
相对定量
animal-derived components
specific primers
cycle threshold value
amplification curve
real-time fluorescent polymerase chain reaction
relative quantification
