蒙成药中“地格达”类原料药材的位点特异性PCR鉴别研究  

Study on identification of "Digeda" raw materials in Mongolian patent medicine by PCR amplification of specific alleles

作　　者：崔占虎[1,2] 黄显章[1] 龙平[2,3] 张乐[3] 赵冬冬[3] 王颖莉[3] 李旻辉[2,3,4] 

机构地区：[1]南阳理工学院,河南南阳473004 [2]道地药材国家重点实验室培育基地中国中医科学院中药资源中心,北京100700 [3]包头医学院,内蒙古包头014060 [4]内蒙古自治区中医药研究所,内蒙古呼和浩特010020

出　　处：《中国中药杂志》2015年第5期793-798,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81060372);国家"十二五"科技支撑计划项目(2012BAI28B02);中医药行业专项(201407003);内蒙古自治区高等学校科学研究项目(NJZY12222)

摘　　要：该文选择8种含有地格达类蒙药原料药材的蒙成药为对象,首先对基原植物肋柱花和苦地丁样品采用改良CTAB法提取其总DNA,使用psb A-trn H片段进行扩增、测序,与Gen Bank下载的8种蒙成药中其他原料药材psb A-trn H序列进行同源比对后根据其变异位点设计特异性鉴别引物。同时对8种蒙成药进行总DNA的提取,并使用DNA纯化试剂盒进行纯化,通过PCR反应对叶绿体rbc L序列进行扩增,再分别选择肋柱花和苦地丁鉴别引物进行扩增,并将扩增后的产物进行测序。所得序列校正、比对后,进行比对分析。结果表明,地格达-4汤、地格达-8散、地格达-4散中均含有原料药材肋柱花,苦地丁特异鉴别引物扩增分析可以鉴定出利胆八味散、伊赫哈日-12和阿嘎日-35中含有苦地丁。该研究结果说明,位点特异PCR方法用于鉴定蒙成药中原料药材具有一定的可行性。To explore a new method for identification of Mongolian patent medicine （MPM） by PCR amplification of specific al- leles. Eight kinds of MPM were used to study the identification of ＂Digeda＂ raw materials. The total DNA of Loma^gonium 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 puri- fication 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 Digeda4 Tang, Digeda-8 San, Digeda4 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 dis- tingnish raw medicinal materials in MPM.

关 键 词：分子鉴别 蒙成药 “地格达” 位点特异性PCR 

分 类 号：R29[医药卫生—民族医学]