基于荧光测序分型技术的六味地黄丸原料分子鉴别研究

该文选择成药六味地黄丸为研究对象,建立了一种用于中成药原料药材基原鉴定的荧光测序分型技术。首先以原料药材丹皮DNA为模板,筛选最适荧光标记鉴别引物,即通过聚合酶链式反应(polymerase chain reaction,PCR)分别对5个FAM荧光标记引物psb A-trn H,ITS,trn L-trn F,mat K,rbc L序列进行扩增,并对扩增产物进行测序,所得测序结果使用GeneMarker V1.80进行分析。根据分析结果最终选择psb A-trn H荧光标记引物来鉴别六味地黄丸及其原料药材。结果表明,通过psb A-trn H荧光标记引物扩增分析,可以准确鉴定出六味地黄丸中的丹皮,山药,泽泻3种原料药材。研究结果说明,采用分子荧光测序分型技术鉴定中成药原料具有一定的可行性。

Study on fluorescence sequencing typing technology identification of raw materials in Liuwei Dihuang pill

In this paper, Liuwei Dihuang pill was used to study the identification of Chinese patent medicine by fluorescence sequencing typing technology. The DNA of Paeonia suffruticosa was used as template to amplify by five pair of FAM fluorescence labeling primers. Then, the amplified products were sequenced. The sequencing results were analyzed by GeneMarker V1.80 to screen the best fluorescence labeling primers. As a result, psbA-trnH fluorescence labeling primer was used to identify the raw materials of Liuwei Dihuang pill. The results showed that three kinds of raw plant medicinal materials in Liuwei Dihuang pill were able to be correctly identified by psbA-trnH fluorescence labeling primer. The fluorescence sequencing typing technology can stably and accurately distinguish raw medicinal materials in Chinese patent medicine.