基于DNA测序方法检测生鲜肉中5种动物源性成分

Identification of 5 kinds of animal-derived components in meat by DNA sequencing

中文关键词:  线粒体  测序  肉类掺假  系统进化树  动物源性成分

英文关键词:mitochondrion  sequencing  meat adulteration  phylogenetic trees  animal-derived components

中文摘要:

      目的 建立生鲜肉中猪、牛、羊、鸡、鸭源性成分的DNA测序鉴别方法, 并对采集自北京各地区生鲜肉品进行检测验证。方法 合成动物线粒体上12S rRNA、cytochrome b、cytochrome c oxidase基因引物, 建立猪、牛、羊、鸡、鸭源性成分的DNA测序鉴别方法, 并对50份猪、牛、羊、鸡、鸭肉品进行检测。结果 使用线粒体上12S rRNA、cytochrome b、cytochrome c oxidase基因引物对猪、牛、羊、鸡、鸭肉品进行扩增, 分别获得456、400和710 bp大小的DNA片段。只有12S rRNA基因引物均可对此5种动物源性成分扩增且效果良好。扩增产物进行序列测定。根据序列构建的系统进化树发现, 此方法可有效区分样品中猪、牛、羊、鸡、鸭源性成分。结论 此方法快速、简便, 可准确检测50种生鲜肉样品中的动物源性成分, 作为肉类鉴别的新方法。

英文摘要:

      Objective To establish the DNA sequencing method for identification of pork, cattle, sheep, chicken and duck in raw meat, and to test and verify the fresh meat purchased from Beijing regions. Methods The primers of 12S rRNA gene, cytochrome b gene and cytochrome c oxidase gene were synthesized, DNA sequencing and identification methods of pork, cattle, sheep, chicken and duck origin were established, and 50 kinds of meat samples of pork, cattle, sheep, chicken and duck were tested. Results The primers of 12S rRNA gene, cytochrome b gene and cytochrome c oxidase gene on mitochondria were used to amplify meat samples of pork, cattle, sheep, chicken and duck, and 456、400 and 710 bp DNA fragments were obtained, respectively. Only primers on 12S rRNA genes could amplify the 5 kinds of animal-derived components, and the effect was good. Amplification products were sequenced. According to phylogenetic tree constructed by the sequence, this method could effectively distinguish the source components of pork, cattle, sheep, chicken and duck. Conclusion The established method is rapid, simple and accurate for the detection of animal-derived components in 50 kinds of raw meat samples, and can be used as a new method to identify meat species.