基于实时荧光PCR对肉制品中羊肉的精确定量

Accurate quantification of mutton in meat products by real-time quantitative PCR

该研究将实时荧光定量PCR技术(real-time fluorescence quantitative PCR,rPCR)与微滴式数字PCR技术(micro-droplet digital PCR,ddPCR)相结合,利用ddPCR建立拷贝数和羊肉质量的函数关系,确立了基于rPCR定量检测羊肉含量的方法。特异性实验结果表明,除绵羊和山羊外,非目标物种DNA未出现特异性扩增;灵敏度实验结果表明,该方法的最低检出限为0.01 ng/μL;通过对已知成分的混合样品和市售样品的检测表明,该方法能够对含量在5%以上的羊肉进行准确定量。因此,该方法在肉制品中羊肉含量检测和掺假鉴别方面具有较好应用潜力。

In order to accurately quantify the content of mutton in meat products,this study established the functional relationship between copy number and mutton weight by using micro-droplet digital PCR(ddPCR).And a quantitative detection method of mutton content based on real-time fluorescence quantitative PCR(rPCR)were also developed.The result of specific experiment showed that the DNA of sheep and goat could be specifically amplified and the detection limit was 0.01 ng/μL.For the mixed samples and the commercial samples,this method could precisely quantify more than 5%of mutton contents in meat products.This method has a potential application in the detection of mutton content in meat products.