[Study on identification of "Digeda" raw materials in Mongolian patent medicine by PCR amplification of specific alleles].
[Study on identification of "Digeda" raw materials in Mongolian patent medicine by PCR amplification of specific alleles].
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DOI：
10.4268/cjcmm20150504
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发表时间：
2015-03
期刊：
影响因子：
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通讯作者：
Zhan-hu Cui;Xian-zhang Huang;P. Long;Le Zhang;Dong-dong Zhao;Ying-li Wang;Minhui Li
中科院分区：
文献类型：
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作者：
Zhan-hu Cui;Xian-zhang Huang;P. Long;Le Zhang;Dong-dong Zhao;Ying-li Wang;Minhui Li
目的探索一种特异性等位基因PCR扩增鉴定蒙药的新方法。采用8种MPM对“地格达”药材进行了鉴别研究。采用改良的CTAB法提取肋柱花和苦杏仁的总DNA，PCR扩增psbA-trnH序列，并进行定向测序。设计特异性引物。用商品化的DNA纯化试剂盒对8种MPM的DNA进行了提取和纯化。用rbcL和两对特异性引物进行扩增。对特异性扩增产物进行正向测序。比对并分析所有特定序列。结果表明，地疙瘩-4汤、地疙瘩-8散、地疙瘩-4散、地疙瘩-4散和C.用利胆八味散、伊和哈日-12和阿加日-35的特异性引物可鉴别出苦皮藤药材。PCR扩增特异等位基因可稳定、准确地区分MPM中的原料药材。
To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.