6种成药中原料药材金银花的分子鉴别研究

目的：研究位点特异性PCR方法在中成药鉴定方面的应用。方法：本实验选用6种含有金银花原料药材的中成药为研究对象,采用改良CTAB法提取其总DNA,并使用DNA纯化试剂盒进行纯化;通过聚合酶链式反应（polymerase chain reaction,PCR）对trnL-trnF叶绿体序列进行扩增,将扩增产物进行不同浓度稀释;以稀释后样品作为模板,分别选择金银花特异性鉴别引物进行扩增,并将扩增后的产物进行测序。所得序列校正、比对后,进行Blast N比对分析。结果：连翘败毒丸、梔子金花丸、牛黄清宫丸、金噪散结丸、健脑补肾丸、金噪开音丸6种中成药中均含有原料药材金银花。结论：采用位点特异PCR方法鉴定中成药中原料药材具有一定的可行性。

Molecular Identification of Raw Material Honeysuckle from Six Kinds of Chinese Patent Medicine

Objective： To study the application of allele specific PCR in identification of Chinese patent medicine. Methods： DNA was extracted by modified CTAB method from 6 kinds of Chinese patent medicine,and was purified by DNA purification kits. The trnL- trnF sequences were amplified,and the amplification products were diluted into different concentration. The dilution samples were used as template,and were amplified by specific primers. The amplified products were sequenced in forward directions. All sequences were aligned and were analyzed by Blast N. Results： Honeysuckle can be identified by honeysuckle primers from Lian Qiao Bai Du Wan,Zhi Zi Jin Hua Wan,Niu Huang Qing Gong Wan,Jin Sang San Jie Wan,Jian Nao Bu Shen Wan,and Jin Sang Kai Yin Wan. Conclusion： PCR amplification of specific alleles can stably and accurately distinguish multi- origin medicinal materials in Chinese patent medicine.