【摘要】 目的:探究多重PCR技术在食品中4种(猪肉、牛肉、羊肉和鸡肉)肉类成分快速鉴别中的应用。方法:采用DNeasy试剂盒提取法、SDS-蛋白酶K法与CTAB-蛋白酶K法提取肉类中总DNA,比较三种方法的效率及纯度,确定最优方式。在动物线粒体细胞色素b基因的差异性位点,设计两套多重PCR引物。建立并优化多重PCR反应体系,根据电泳检测扩增产物分子量差异以快速鉴别4种肉类。结果:DNeasy试剂盒法、SDS-蛋白酶K法提取DNA效率显著优于CTAB-蛋白酶K法,P<0.05。在优化反应条件下选择特异性高、序列长的多重PCR引物可有效快速鉴定食品中猪、牛、羊与鸡源性成分,检测灵敏度为皮克级。结论:多重PCR方法应用于食物中多动物源性成分鉴别,精确稳定快速。更多还原

【Abstract】 Objective: To explore the rapid identification for 4 kinds of meat(pork, beef, mutton and chicken) in food by multiplex PCR technique. Methods: The total DNA was extracted by DNeasy kit extraction, SDS-protease K and CTAB-protease K; the efficiency and purity of the 3 methods were compared; then the optimal method was determined. 2 sets of multiplex PCR primers were designed in the mitochondrial cytochrome b gene of animals. The multiplex PCR reaction system were established and optimized, to rapidly identify 4 kinds of meat according to the difference of molecular weight by electrophoresis. Results: The extraction efficiency of DNeasy kit method, SDS-protease K is significantly better than the CTAB-protease K method, with statistical difference, P<0.05. On the optimal reaction conditions that high specificity, sequence length of the multiplex PCR primers can be effectively and rapidly identify food ingredients of pigs, cattle, sheep and chicken; the detection sensitivity is peake. Conclusion: Application of multiplex PCR technology in identifying meat is accurate, stable and quick.更多还原