应用PCR-RFLP方法鉴定梅花鹿茸与马鹿茸

An Experimental Study on the Identification of Sika Deer Antler and Cervus Pilose Antler by PCR-RFLP

目的建立一种应用限制性片段长度多态性技术鉴别梅花鹿茸与马鹿茸的方法。方法提取各种鹿茸样品的基因组DNA,通过聚合酶链式反应(polymerase chain reaction,PCR)对正品鹿茸的特异性片段进行扩增,鉴别鹿茸的真伪,然后对正品鹿茸利用动物通用引物进行PCR扩增,并通过限制性片段长度多态性分析进一步对正品鹿茸的种属来源进行确证。结果通过对特异性片段的PCR扩增,可将梅花鹿茸及马鹿茸与驯鹿、麋鹿、新西兰鹿等伪品鹿茸加以区分,通过对限制性内切酶Msp I进行酶切后的片段长度进行分析,可进一步区分梅花鹿茸与马鹿茸。结论本实验建立了一种基于限制性片段长度多态性技术快速、准确的鉴别鹿茸真伪及种属鉴定的方法,为解决中药易混品种难以鉴定的问题提供了又一个新方法。

OBJECTIVE To establish a method of restriction fragment length polymorphism(RFLP) to distinguish sika deer antler and cervus pilose antler. METHODS The genomic DNA of various samples of pilose antler was extracted, the specific fragments of the genuine pilose antler were amplified by polymerase chain reaction(PCR) to identify the true and false of pilose antler, and then the genuine pilose antler was amplified by PCR using universal primers of animals, and the provenance of the genuine pilose antler was further confirmed by restriction fragment length polymorphism analysis. RESULTS By PCR amplification of specific fragments, we can distinguish the antler of sika deer and cervus pilosula from those of reindeer, elk and New Zealand deer. By analyzing the fragment length after restriction endonuclease Msp I, we can further distinguish the antler of sika deer and cervus pilosula. CONCLUSION This study established a rapid and accurate identification method of velvet antler and species identification based on restriction fragment length polymorphism technology, which provides another new method to solve the problem of identification of easily mixed varieties of traditional Chinese medicine.