应用实时荧光PCR技术量化检测羊肉中猪源性和鸡源性成份

Detection and quantification of pork and chicken in sheep products using real-time PCR

中文关键词:  荧光实时定量PCR  定量检测  食品掺假  羊肉  猪肉  鸡肉

英文关键词:real-time PCR  quantitative analysis  food adulteration  sheep  pork  chicken

中文摘要:

      目的 建立羊肉中猪源性成份和鸡源性成份的定量检测方法。方法 以新鲜羊、猪和鸡瘦肉为样本提取DNA分子作为检测模板, 针对基因组中单拷贝基因设计特异性的引物和探针, 应用荧光实时定量PCR技术对模板DNA进行扩增。通过绘制扩增标准曲线和确定羊、猪和鸡的质量与DNA比值常数, 对4种不同掺混比例的混合肉样进行定量分析。结果 检测质量百分比的绝对误差可以控制在7%以内, 量化结果基本准确。结论 对于组织成份单一的样品, 可以通过在基因组单拷贝基因上设计特异性的引物, 利用PCR技术实现在质量水平上对食品中动物源性成份的量化分析, 该技术方法的建立可以为肉类掺假的监管工作提供有力的技术支撑。

英文摘要:

      Objective To establish a method to determine pork and chicken adulteration in mutton. Methods A series of genome DNA were extracted from fresh pork and chicken as detection template, and the primers were designed according to the sequence of the single copy gene. Amplification of detection template were conducted through real-time fluorescent quantitative PCR technique. The mathematical conversion parameters about the mass of meat and DNA were determined by standard curve. The weight/weight equivalents of 4 blending samples were analyzed. Results The results showed that the absolute error of detection with a tolerance was less than 7%, and the quantitative results were accurate. Conclusion Quantitative analyzing of the composition of the animal species in single component sample can be realized with real-time PCR by designing the primer according to the sequence of genome. The current research can provide technical support for food safety detection.