基于DNA条形码技术对朝鲜牛黄安宫丸中犀牛角成分鉴别的研究

Research on the Species Identification of Rhinoceros Horn in Bezoar Angong pill of Korea based on DNA Bar-coding Technique

目的建立DNA条形码技术对朝鲜牛黄安宫丸中犀牛角成分的真伪鉴别方法,对打击非法贸易及保护珍稀濒危野生动物具有重大意义.方法以涉案朝鲜牛黄安宫丸为材料,根据材料自身的特点对其进行消化处理后采用市售动物基因组核酸提取试剂盒方法提取核酸,并采用DNA条形码片段12S rRNA通用引物对提取的基因组DNA进行PCR扩增反应和测序.结果对测序结果使用NCBI中Blast进行序列相似性分析比对,结果表明5份涉案朝鲜牛黄安宫丸的扩增产物序列与非洲水牛的线粒体DNA的12S rRNA基因序列的部分片段的一致性达98.84%,因此为水牛角成分并非使用濒危物种犀牛角.结论依靠分子生物学中DNA条形码技术对疑似犀牛角药材进行准确的物种鉴定,可用于鉴别犀牛角药材的真伪,为各执法部门提供可靠的技术支持,规范国内和口岸犀牛角制品市场.

Objective To establish the rhinoceros horn in Bezoar Angong pill of Korea authenticity methods based on DNA bar-coding technique,it will help to combat illegal trade and protect rare and endangered wild animals.Methods Use the case involved Bezoar Angong pill of Korea as materials,according to the characteristics of material used for the digestion treatment,the kit for extraction of nucleic acid from animal genome in market was used to extract the total DNA,then the Genomic DNA was amplified and sequenced by using 12 S rRNA universal primers of DNA bar-code fragment.Results The similarity alignment and analysis of sequences with NCBI in Blast,and then the phylogenetic tree was built.The results show that the obtained sequences from Bezoar Angong pill of Korea show 98.84%consistent with partial mitochrondrial DNA 12S rRNA gene of Synceruscaffer.So it is made from Buffalo Horn instead of the endangered species Rhino Horn.Conclusion The species of rhinoceros horn medicinal materials could be accurate determined by using DNA bar-coding technique of molecular biology.It can be used to identify the authenticity of rhinoceros horn medicinal materials,offer reliable technical support for law enforcement agencies and regulate the domestic and port rhinoceros Horn products market.