荧光定量PCR方法检测畜肉食品中猪源性成分

Detection of porcine-derived materials in meat products by real time PCR method

目的建立一种快速、特异、灵敏的猪源性成分检测方法。方法本研究以猪线粒体12S rRNA基因序列为靶位点设计引物和探针,进行荧光定量PCR扩增,建立猪源性成分检测方法;以常见畜禽肉包括羊肉、牛肉、鸡肉、鹅肉、鸭肉、兔肉、马肉、鹿肉等参考动物物种作特异性检测;以50 mg/kg羊肉DNA作为稀释液对猪肉DNA进行梯度稀释,做灵敏度检测。结果该方法能够有效对猪源性成分进行快速检测,具有较强的特异性,灵敏度较高(可达0.1μg/kg)并且羊肉成分的存在对猪肉灵敏度检测没有影响。结论该方法特异性强,灵敏度高,可以快速、准确检测畜肉食品中含有的猪源性成分。

Objective To establish a rapid, specific, sensitive detection method for porcine derived mate-rials. Methods A real-time PCR method for the porcine derived materials detection in meat products was es-tablished using forward and reverse primers and probe corresponding to the mitochondrial 12S rRNA gene. The specific detection was carried out by livestock and poultry meat including lamb, beef, chicken, goose, duck, rabbit meat, horse meat, venison and other reference animal species and 50 mg/kg lamb DNA was chosen as diluent by gradient dilution of pork DNA, then sensitivity detection was executed. Results This method could effectively detect the porcine-derived materials, and showed a strong specificity and high sensitivity (up to 0.1μg/kg), and the mutton components had no obvious influence on pork sensitivity detection. Conclusion This method could detect the porcine-derived components in meat products rapidly and accurately.