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[采用等位基因特异性PCR对金银花配方颗粒进行分子鉴定]

[Molecular authentication of Jinyinhua formula granule by using allele-specific PCR].

作者信息

Jiang Chao, Tu Li-Chan, Yuan Yuan, Huang Lu-Qi, Gao Wei, Jin Yan

机构信息

State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.

School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2017 Jul;42(13):2484-2491. doi: 10.19540/j.cnki.cjcmm.20170527.002.

Abstract

Traditional authentication method is hard to identify herb's authenticity of traditional Chinese medicine(TCM) formula granules because they have lost all their morphological characteristics. In this study, a new allele-specific PCR method was established for identifying the authentication of Jinyinhua formula granule (made from Lonicerae Japonicae Flos) based on an SNP site in trnL-trnF fragment. Genomic DNA was successfully extracted from Lonicerae Japonicae Flos and its formula granules by using an improved spin column method and then PCR was performed with the designed primer. Approximately 110 bp specific bands was obtained only in the authentic Lonicerae Japonicae Flos and its formula granules, while no bands were found in fake mixed products. In addition, the PCR product sequence was proved from Lonicerae Japonicae Flos trnL-trnF sequence by using BLAST method. Therefore, DNA molecular authentication method could make up the limitations of character identification method and microscopic identification, and quickly identify herb's authenticity of TCM formula granules, with enormous potential for market supervision and quality control.

摘要

传统的鉴定方法难以鉴别中药配方颗粒中药材的真伪,因为它们已失去了所有的形态特征。在本研究中,基于trnL-trnF片段中的一个单核苷酸多态性(SNP)位点,建立了一种新的等位基因特异性PCR方法,用于鉴别金银花配方颗粒(由金银花制成)的真伪。采用改进的离心柱法成功从金银花及其配方颗粒中提取了基因组DNA,然后用设计的引物进行PCR。仅在正品金银花及其配方颗粒中获得了约110 bp的特异性条带,而在假冒混合产品中未发现条带。此外,通过BLAST方法从金银花trnL-trnF序列中验证了PCR产物序列。因此,DNA分子鉴定方法可以弥补性状鉴定方法和显微鉴定的局限性,并能快速鉴别中药配方颗粒中药材的真伪,在市场监管和质量控制方面具有巨大潜力。

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