应用二重PCR技术检测肉制品中的肉源性成分

Detection of Meat-derived Components in Meat Products by Diplex PCR

在建立单重PCR的基础上成功建立了多种二重PCR体系,以同时检测2种肉源成分(猪、牛、鸡和羊)。PCR扩增的是每种动物的线粒体基因,由于鸡、羊的PCR扩增片段大小过于接近(131和119 bp),电泳图上不易区分,故试验中的二重PCR体系使用了5种动物组合。所建立的二重PCR方法能够迅速、稳定地同时鉴别2种肉源性成分,为下一步建立三重或四重PCR方法奠定了良好的基础。

In the current study, multiple diplex PCR systems were developed in order to simultaneously detect two animal DNA derived from pork,beef,sheep or chicken. The diplex PCR systems were constructed based on previously designed single PCRs using mitochondrial genes of each animal species. The combination of chicken with sheep was not used in the study since the amplicon sizes of these two animal species were too closely to distinguish(131 and 119 bp). Therefore,the diplex PCRs system used in the experiment used five animal combinations. The diplex PCR procedure standardised in this study is simple,reliable and efficient to detect meat adulteration containing pork,beef,chicken and sheep. The results can also be used to develop multiplex PCR containing three or four of these animal species. The diplex PCR procedure standardised in this study is simple. The results could also be used to develop multiplex PCR containing three or four of these animal species.