Molecular Identification of Chinese Patent Medicine Containing Zaocys dhumnades Based on 12S Gene
OBJECTIVE To establish a molecular identification method for the adulteration of Zaocys dhumnades in Chinese patent medicine.METHODS Eleven kinds of Chinese patent medicines containing Zaocys dhumnades and 15 kinds of Zaocys dhum-nades and their adulterants were collected.The total genomic DNA was extracted from all samples,and the comparison and analysis of Zaocys dhumnades and its adulterants were conducted based on 12S gene fragments.The universal primer sequence,the specific primer sequence and the primers and probes used for fluorescence quantification of Zaocys dhumnades were designed according to the differ-ence points.Different PCR methods were used to amplify the DNA of Zaocys dhumnades contained in the medicinal materials of Zaocys dhumnades and Chinese patent medicines respectively.The reaction system was optimized and the tolerance and adaptability of the methods were investigated.RESULTS The specific PCR technique of Zaocys dhumnades was established.All the authentic Zaocys dhumnades had a clear band around 200 bp,and other adulterants had no band.Fluorescent quantitative PCR method was established for the identification of Chinese patent medicines with low DNA content CONCLUSION The medicinal materials of Zaocys dhum-nades can be effectively identified by PCR amplification with specific primers.Fluorescent quantitative PCR detection can effectively identify the authenticity of Zaocys dhumnades in Chinese traditional medicine.The method is specific,rapid and sensitive,which provides a reference for the identification of Zaocys dhumnades in Chinese patent medicine.
NETL
NSTL
万方数据
