利用多重等位基因特异PCR鉴别人参、西洋参

Application of multiplex allele-specific PCR for authentication of Panax Ginseng and P.quinquefolius

目的:查找并发现参类药材的SNP位点,利用多重等位基因特异PCR同时鉴别人参、西洋参。方法:查找GenBank上已收载的参类药材序列,进行比对分析,根据人参、西洋参的SNP位点设计引物,对PCR反应体系进行优选。在此基础上,对20个不同来源的人参、西洋参样品进行PCR扩增,根据各自的特异条带进行鉴别。结果:当退火温度为66℃时,出现249 bp条带的为人参,1 049 bp条带的为西洋参。该鉴别反应特异性高、重复性好,能在同一反应中准确鉴别人参、西洋参。结论:多重等位基因特异PCR能成功地对人参、西洋参进行鉴别,在中药材分子鉴定中具有推广应用价值。

Objective： Searching and identifying SNP in Panax species and using multiplex allele-specific PCR （ MAS-PCR）to authenticate P. ginseng and P. quenquefolium. Method ： Based on genbank database of Panax species, using DNAMAN to align the sequences, identify SNP of P. ginseng and P. quenquefolium. Design allele-specific primers for P. ginseng and P. quenquefolium, optimize the PCR reaction system including the usage amount of Taq, dNTP, primer, etc. Optimized system was performed with the total DNA of 20 different sources of P. ginseng and P. quenquefolium. Result： When the annealing temperature was 66 ℃, the template DNA of P. ginseng could be amplified 249 bp band whereas P. quenquefolium amplified 1 049 bp band. Conclusion： The MAS-PCR have the advantages of highly specific, good reproducibility and could be identify P. ginseng and P. quenquefolium in the same PCR tube. It was a potential method to use in the molecular identification of other meteria medica.