基于TaqMan实时荧光PCR检测鲜肉及加工肉制品中的驴源性成分

根据出入境检验检疫行业标准SN/T 3730.4—2013《食品及饲料中常见畜类品种的鉴定方法 第4部分：驴 成分检测 实时荧光PCR法》合成引物和探针，利用TaqMan实时荧光聚合酶链式反应（polymerase chain reaction， PCR）技术检测鲜肉及加工肉制品中的驴源性成分。首先对13 种不同动物鲜肉组织的DNA进行驴源性成分特异 性检测，然后对驴源性DNA模板原液进行梯度稀释，检测方法灵敏度，最后在加工肉制品中检测方法的适用性。 结果表明：本研究建立的方法特异性强，除驴肉外，牛、羊、猪、马、骆驼、鹿、狗、兔、鸡、鸭、鸽子、鹌鹑 12 种动物鲜肉组织均无特异性扩增；方法的灵敏度较高，驴组分DNA的检出限可达100 fg/μL，灵敏度可达0.01%； 方法的适用性较广，可以用于加工肉制品中驴源性成分的检测。

Identification of Donkey-Derived Ingredients in Raw and Processed Meats Using TaqMan Probe Based Real-Time Polymerase Chain Reaction

In this study, we synthesized a set of primers and TaqMan probe according to the industry standard (SN/T 3730.4—2013) for the detection of donkey-derived ingredients in raw and processed meats by real-time fluorescence polymerase chain reaction (PCR). The specificity of the primers and probe were tested using DNA templates extracted from raw meats from donkey and 12 other animal species. Gradient dilutions of the donkey-derived DNA template were used to evaluate the sensitivity. The applicability of the PCR assay for processed meat was also investigated. The results showed that this PCR assay was highly specific and could specifically amplify DNA from donkey rather than cattle, sheep, pig, horse, camel, deer, dog, rabbit, chicken, duck, pigeon and quail. It was sensitive. The limit of detection for donkey DNA was 100 fg/μL and the sensitivity was 0.01%. This method was widely applicable and useful for the detection of donkey-derived ingredients in processed meat.