实时定量PCR法对牛肉中鸡源性成份的量化检测

Real-time PCR method for quantitative detection of chicken blending in beef

中文关键词:  肉类掺假监管  量化研究  实时荧光定量PCR  鸡肉  牛肉

英文关键词:meat adulteration  quantitative analysis  real-time PCR  chicken  beef

中文摘要:

      目的 实现样品中牛源性成份和鸡源性成份的量化分析。方法 通过在基因组单拷贝基因上设计引物, 绘制模板DNA扩增标准曲线以及确定牛肉、鸡肉质量与DNA浓度的比值常数, 利用实时荧光定量PCR技术对四种不同掺混比例的牛鸡瘦肉混合样本中牛源性成份和鸡源性成份所占的质量百分比含量进行分析。结果 通过荧光实时定量PCR反应的Ct值、模板DNA扩增标准曲线和质量与DNA的比值常数可以计算出样品中所含牛源性成份和鸡源性成份的质量百分比含量, 检测值与理论值之间的绝对误差可控制在5%以内, 量化研究结果基本准确。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监管工作提供有力的技术支撑。

英文摘要:

      Objective To quantitatively analyze the beef and chicken in blending samples. Methods The primers were designed according to the sequence of the single copy gene. A series of genome DNA were used as standard curve in real-time PCR. The mathematical conversion parameters about the mass of meat and DNA were determined. The weight/weight equivalents of beef and chicken in four blending samples were analyzed using real-time PCR. Results The weight/weight equivalents of beef and chicken was determined by calculated using Ct, standard curve and the mathematical conversion parameters about the mass of meat and DNA. The results showed that the absolute error of detection with a tolerance of less than 5%. Conclusion Quantitative analyzing of the composition of the animal species in single component sample could be realized using real-time PCR by designing the primer according to the sequence of genome. This research could provide technical support for food safety.