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目的常用生药柴胡正品为柴胡属植物柴胡及狭叶柴胡的干燥根。当前,同属多种植物也做柴胡内转录间隔区使用。仅凭生药性状及显微特征难以进行真伪鉴别。在本实验中,笔者针对6种不同柴胡属植物来源的柴胡生药,基于其内转录间隔区(ITS1)区多态性研制了用于其鉴别的DNA芯片。方法将设计的特异性寡核苷酸探针点印于基质上制备芯片;从待检测样品中提取基因组DNA,扩增ITS1片段,并以荧光标记后,与芯片杂交。结果在与其相应的探针位置即可出现信号;每种不同来源的柴胡有其特异的杂交谱图。结论研制的DNA芯片可为柴胡类生药的鉴定提供可靠便利的途径。
Purpose The common crude drug Bupleurum is the dry root of Bupleurum and Bivalvia. At present, the same species of Bupleurum are also used within the transcribed spacer. It is difficult to authenticate the authenticity only by the properties of crude drugs and microscopic features. In this experiment, the author for the six different Bupleurum origin of Bupleurum, based on its internal transcriptional interval (ITS1) region polymorphism developed for the identification of DNA chips. Methods The specific oligonucleotide probes were designed and printed on the substrate to prepare the chip. The genomic DNA was extracted from the sample to be tested, ITS1 fragment was amplified, labeled with fluorescent and then hybridized with the chip. As a result, a signal appears at the position of the corresponding probe; each different source of Bupleurum has its own specific hybridization pattern. Conclusion The developed DNA chip can provide a reliable and convenient way for the identification of Bupleurum crude drugs.