Identification of Salvia miltiorrhiza and Its Adulterants by High-Resolution Melting Analysis
The objective of this study was to establish a new method for identifying Salvia miltiorrhiza and its adulterants effectively based on high-resolution melting(HRM)technology.In this study,the genomic DNA was purified by silica gel column and specific primers were designed based on the rDNA ITS2 sequence of S.officinalis.The melting curve model of ITS2 was established by systematically optimizing the concentra-tions of primer and DNA template,annealing temperature and cycle times,and the HRM analysis was used to identify S.miltiorrhiza and its adulterants(S.przewalskii,S.yunnanensis and S.officinalis).The results showed that the amplification was observed to be stable,uniform and specific under the optimal concentrations of primer and template of 2.5 to 10.0 μmol/L and 26.95 to 53.90 ng/μL,the annealing temperature of 58 ℃,and the cycle number of 35 to 45,respectively.Through comparative analysis of melting temperature(Tm),it was achievable to identify S.miltiorrhiza from S.przewalskii,S.yunnanensis,and S.offiicinalis,showing u-nique advantages in rapid detection of S.miltiorrhiza and its adulterants.
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