微滴式数字PCR与实时荧光PCR检测羊肉制品中羊源和猪源性成分方法的比较

Comparison of microsphere digital PCR and real-time fluorescence PCR for detection mutton-derived and porcine-derived ingredients in mutton products

参考外文文献中羊肉和猪肉的特异性引物及Taq Man探针,分别建立一种定量检测羊肉制品中羊源和猪源性成分的微滴式数字PCR方法,并将该方法与标准SN/T 2051-2008中实时荧光PCR方法做对比来检测3份羊肉制品中的羊源和猪源性成分。结果表明,实时荧光PCR方法检测5号、9号、23号样品中的羊源性成分的Ct值分别是14.424/19.214、18.345/20.147、17.321/20.542;猪源性成分的Ct值分别是18.923/34.483、20.121/28.963、20.352/33.851;微滴式数字PCR方法检测5号、9号、23号样品中的羊源和猪源性成分的DNA拷贝数分别为236/0 copies/μL、421/0.3copies/μL、402/0.11copies/μL。表明两种方法均能检出3份样品中均含有羊源和猪源性成分,而微滴式数字PCR方法能够对DNA模板定量分析。结论:在肉种成分真伪鉴定上,微滴式数字PCR方法较实时荧光PCR方法更科学、准确。

The specific primers and TaqMan probes in foreign literatures were referred. The micro- sphere digital PCR method for quantitative detection of mutton - derived and porcine - derived ingredients in mutton products was established. The method was compared with real -time fluorescence PCR method in SN/T 2051 -2008 to detect the mutton - derived and porcine - derived ingredients of three parts of mutton products. The results showed that the Ct values of No. 5, No. 9 and No. 23 samples were 14. 424/19. 214,18. 345/20. 147,17. 321/20. 542 respectively for mutton - derived ingredients and 18. 923/34. 483,20. 121/28. 963,20. 352/33. 851 respectively for porcine - derived ingredients by the fluorescence PCR method. And the DNA copies of mutton - derived and porcine - derived ingredients in the three samples were 236/0 copies/μL,421/0.3 copies/μL,402/0.11 copies/μL detected by the mi- erosphere digital PCR method. It showed that both the fluoreseenee PCR method and the microsphere digital PCR method could detect mutton - derived and porcine - derived ingredients in the three samples. But the microsphere digital PCR method could do quantitative analysis to the DNA templates. The conclusion was that for identifying the authenticity of ingredients in meat products, the microsphere digital PCR method was much more scientific and accurate than the fluorescence PCR method.