PCR方法特异性鉴定狮源性制品

Study on the PCR Method for Specific Identification of Panthera Leo

目的建立可对狮DNA进行特异性检测的PCR方法,应用在检验检疫过程中获得的狮疑似样品和保护执法工作中难以检查辨认的狮产品进行物种鉴定。方法针对狮属动物线粒体细胞色素b(Cytb)基因设计特异性引物,用该引物分别对从亚洲狮、非洲狮和虎、豹、熊等14种非狮源性哺乳动物的肌肉、血液、毛发和肉骨粉中提取的DNA进行PCR(聚合酶链反应)扩增。结果试验表明所设计的引物对狮DNA具有特异性,从而达到了对该物种进行特异性检测鉴定的目的。检测灵敏度可达到在骨粉中检出0.5%含量的狮源性成分。结论该PCR方法具有方便、经济、灵敏度高和重复好等特性,可用于狮源性制品的检测鉴定。

Objective To develop a PCR for specific detection of DNA from Panthera Leo species, for Panthera Leo species identification of suspected samples or unrecognized products obtained in quarantine investigation or forensic tests. Method A pair of primers was designed specific to the cytochrome b gene in D-loop DNA fragment of mitochondria. PCR tests with the primer were performed to amplify DNAs extracted from muscle,blood, hair or bone meal of different P.leo breeds and 14 mammalian species other than P. leo. Results The results indicated that the primers were specific to Panthera leo DNA and Panthera Leo species could be accurately identified. The method was sensitive,reproducible and reliable. Conclusion The method could be applied to food detection, fur identification and traditional Chinese drug identification.