基于ITS2序列位点特异性PCR鉴别广藿香及其混伪品

Identification of Pogostemon cablin (Blanco) Benth. with the Counterfeit Products by Allele-specific PCR Based on ITS2 Sequence

目的建立一种简便、准确鉴别广藿香及其混伪品的方法。方法通过对GenBank数据库收录的广藿香及其混伪品的ITS2序列进行对比分析,根据差异位点设计位点特异性鉴别引物,并优化PCR条件,对广藿香新鲜植物、干燥药材、含广藿香中成药及其混伪品进行扩增和检测。结果广藿香新鲜植物、干燥药材及含广藿香中成药均扩增出162 bp条带,而广藿香的混伪品均无条带,且该方法灵敏度高,检出限可达0.3 g·L^(-1)。结论所建立的位点特异性PCR方法可实现广藿香的准确鉴别,具有操作简便、稳定可靠、应用范围广的优点。

Objective Designing ARMS specific primer pairs to provide a molecular method for the identification of Pogostemon cablin(Blanco) Benth. and the counterfeit products. Methods Based on the ITS2 genes sequence of P. cablin and its counterfeit species, a pair of allele-specific diagnostic primers were designed to authenticate P. cablin from its counterfeit species. The PCR amplications were performed using the diagnostic primers with the total DNAs of the original plants, dry medicine or Chinese patent medicine as templates. Results Only the template DNA of P. cablin could be amplified whereas the diagnostic PCRs of its counterfeit species were all negative. Conclusion Compared with the other authentification methods, the allele-specific PCR amplification is not only simpler and time-saving but practical and effective.