实时荧光定量PCR方法实现羊肉中鸡成分的精确定量

Real-time Quantitative PCR Method for Accurate Quantification of Chicken Components in Mutton

近年来,世界各地肉制品掺假情况频繁发生。为维护市场稳定,保障食品安全,迫切需要量化肉制品中所含动物成分的质量百分比。本研究建立了一种实时荧光定量PCR定量检测系统,借助微滴式数字PCR技术(droplet digital,ddPCR),有效实现靶向基因拷贝数的绝对定量,利用实时荧光定量PCR技术(real-time quantitative PCR detecting system,q-PCR),将扩增Ct值转化为基因拷贝数,再从基因拷贝数转换为肉的质量,有效实现了Ct值与质量间的转化,从而实现了羊肉中掺假鸡肉的精确定量。实验数据表明,该方法可以很好地应用于掺假羊肉中鸡肉质量百分比的检测,鸡肉的最低定量检测限(LOD)为5%,在5%~80%范围内相对误差小于±9%,相对标准偏差(RSD)小于±20%,定量结果准确、重复性高,可以有效区分生产污染与故意掺假,为肉类掺假的监管工作提供有力的技术参考。

In recent years,adulteration of meat products around the world has occurred frequently.In order to maintain market stability and ensure food safety,it is urgent to quantify the percentage of animal components contained in meat products.In this study,we established a real-time quantitative PCR quantitative detection system,using droplet digital PCR technology(droplet digital,ddPCR)to effectively achieve absolute quantification of target gene copy number,using real-time PCR Real-time Quantitative PCR Detecting System(q-PCR),which converts the amplified Ct value into a gene copy number,and then converts the gene copy number into meat quality,effectively realizing the conversion between Ct value and mass.An accurate quantitative detection method for adulterated chicken in lamb was established.The experimental data show that the method can be applied to the detection of the percentage of chicken meat in mutton.The minimum quantitative detection limit(LOD)of chicken is 5%,and the relative error is less than±9%in the range of 5%to 80%.The deviation(RSD)is less than±20%.The quantitative results are accurate and reproducible.It can effectively distinguish between production pollution and intentional adulteration,and provide a powerful technical reference for the supervision of meat adulteration.