基于实时荧光PCR技术鉴别梅花鹿

Identification of Cervus nippon by fluorescent real-time PCR method

中文关键词:  物种鉴定  梅花鹿  实时荧光PCR  线粒体D-loop基因

英文关键词:species identification  Cervus nippon  real-time PCR  mitochondrial D-loop gene

中文摘要:

      目的 建立基于TaqMan实时荧光PCR技术鉴别梅花鹿的分析方法。方法 通过对梅花鹿线粒体D-loop基因序列比对分析, 选取梅花鹿D-loop基因特异位点作为扩增靶序列, 设计梅花鹿特异性的引物和TaqMan探针, 并用已知样本对引物和TaqMan探针的物种特异性、灵敏性进行验证。结果 该引物和探针对梅花鹿成分检测具有较强的特异性, 与其他物种DNA无交叉扩增, 该方法的检测灵敏度为0.01 ng/μL。结论 本方法具有较高的特异性和灵敏度, 适用于对肉类以及其他梅花鹿产品的物种的鉴别。

英文摘要:

      Objective To establish a method for identification of Cervus nippon based on TaqMan real-time fluorescence PCR. Methods By comparing and analyzing the D-loop gene sequence of Cervus nippon mitochondria, the specific locus of D-loop gene of Cervus nippon was selected as the amplification target sequence, and the specific primers and TaqMan probes of Cervus nippon were designed. The species specificity and sensitivity of the primers and TaqMan probes were verified with known samples. Results The primers and probes showed strong specificity for the detection of Cervus nippon and no cross amplification with DNA of other species. The detection sensitivity of this method was 0.01 ng/L. Conclusion This method has high specificity and sensitivity, and is suitable for identifying the species of meat and other Cervus nippon products.