目的 采用DNA条形码分子鉴定技术鉴别市售柴胡药材及其混伪品，以确保柴胡药材质量及临床用药安全。方法 共收集85份柴胡药材，对其ITS2序列进行PCR扩增并双向测序，所得序列经CodonCode Aligner校对拼接后，利用MEGA 6.0对序列进行分析比对，计算种内、种间遗传距离，利用邻接法（NJ）构建系统聚类树，并应用ITS2数据库网站预测其ITS2二级结构。结果 柴胡种内遗传距离明显小于柴胡与其近缘物种种间遗传距离，基于ITS2建立的NJ树和网站预测的ITS2二级结构，均能将柴胡药材及其混伪品区分开。85份样品中，55份符合《中国药典》2015年版规定的柴胡正品来源，正品率为64.7%。结论 基于ITS2序列可以准确可靠地鉴别柴胡药材及其混伪品，为确保临床用药安全提供新的技术手段。

Objective DNA barcoding technology, a molecular identification method, is applied to distinguishing Bupleuri Radix from its adulterants in order to ensure the quality and clinical curative effect. Methods In this study, the internal transcribed spacer 2 (ITS2) regions of 85 samples were amplified by PCR and sequenced bi-directionally. Obtained sequences were assembled using CodonCode Aligner. The genetic distances were computed by MEGA 6.0 in accordance with the kimura 2-parameter (K2P) model and the phylogenetic tree was constructed by Neighbor-joining (NJ) method. Moreover, the secondary structure of ITS2 was predicted using ITS2 database websites. Results The intra-specific genetic distances were smaller than inter-specific ones in ITS2 regions of Bupleuri Radix. NJ tree and secondary structure results could distinctly differentiate quality product and adulterants. Only 64.7% of the 85 samples were in accordance with the requirements of Chinese Pharmacopoeia. Conclusion ITS2 sequence can accurately and reliably identify the authenticity of Bupleuri Radix and could provide a new technique to ensure clinical safety of this traditional Chinese medicine.

国家自然科学基金资助项目（81673548）；辽宁省重大科技攻关计划（2013226050）；国家中医药管理局项目（201407002）