基于微滴式数字PCR的羊肉中鸭源性成分定量检测

A Quantitative Assay for Measuring Duck Adulterated into Mutton Products by Droplet Digital PCR

为实现羊肉中鸭源性成分定量检测,文章基于微滴式数字PCR技术,建立了能够同时对羊和鸭源性成分定量检测的ddPCR检测方法。根据羊和鸭的单拷贝特异性基因(RPA1)进行引物与探针设计,采用增加PCR循环数等方式对PCR反应条件进行优化。结果表明:通过构建待测样品中鸭肉组分与基因拷贝数的线性关系,模拟混合样本中鸭肉掺杂质量分数10%~90%时,采用ddPCR构建的羊肉和鸭肉标准曲线R;分别为0.995和0.998,预测结果准确;结合市售6批次问题样本与标准样品多重验证表明绝对误差范围在–0.68%到0.35%范围内。综上所述,该方法定量结果准确、重复性高,为食品安全监管工作和相关执法提供了技术保障。

The objective of this work is to establish a method for the quantitative determination of duck-derived components in mutton by droplet digital PCR (ddPCR).The primers and probes were designed according to the single copy specific gene (RPA1) of sheep and ducks.A method for quantitative detection of sheep and duck-derived components by ddPCR was established.The PCR reaction conditions were optimized by increasing the number of PCR cycles to improve the accuracy of detection.At the same time,the linear relationship between the gene copy number and the meat content of duck samples was determined.Results show that when the doping mass fraction of duck meat in the simulated mixed sample is 10%~90%,the R;values of mutton and duck were 0.995 and 0.998,respectively,which were accurate.The absolute error range of standard sample doping quantitative detection was–0.68% to 0.35%,which accurately realized the quantitative detection of duck meat doping in 6 batches of mutton products on the market.In conclusion,the quantitative results of this method are accurate and reproducible,which provides technical support for food safety supervision and related law enforcement.