基于ITS2条形码SNPs的人参和西洋参PCR-SSCP分子鉴别研究

目的:依据人参和西洋参ITS2条形码的专属性SNP鉴别位点,建立利用PCR-SSCP技术鉴别人参和西洋参的方法.方法:查找GenBank上已收录的人参和西洋参ITS2序列,进行比对分析,筛选人参和西洋参ITS2序列的专属性SNP鉴别位点,据此设计引物,采用聚合酶链反应-单链构象多态性(PCR-SSCP)方法对11个人参样品和10个西洋参样品进行分析,对PCR产物进行测序验证.结果:人参和西洋参样品的PCR-SSCP谱带与各自的对照药材谱带一致,人参和西洋参的PCR-SSCP谱带有显著差异；PCR-SSCP鉴别结果和经测序鉴别结果一致.结论:和测序法相比,PCR-SSCP方法快速、准确,可用于人参和西洋参药材的鉴定.

PCR-SSCP molecular identification of Panax ginseng and P. quinquefolius based on ITS2 bar coding SNPs

Objective: To establish a method for identifying Panax ginseng and P. quinquefolius with PCR-SSCP, on the basis of specific SNP identification sites of their ITS2 bar codes. Method: ITS2 sequences of P. ginseng and P. quinquefolius recorded in GenBank were searched to conduct a comparative analysis and screen out specific SNP identification sites of their ITS2 bar codes. Based on that, the Polymerase Chain Reaction-Single Strand Conformation Polymorphism(PCR-SSCP) method was adopted for analyzing 11 P. ginseng samples and 10 P. quinquefolius samples and verifying sequencing of their PCR products. Result: The P. ginseng and P. quinquefolius samples had the same agarose mages of PCR-SSCP with the standard medicines. There were significant differences between the PCR-SSCP agarose mages of P. ginseng and P. quinquefolius, with identifical identification results between PCR-SSCP and sequencing. Conclusion: Compared with the sequencing method, PCR-SSCP is so rapid and accurate that it can be used for identifying P. ginseng and P. quinquefolius medicines.