中国现代中药2025，Vol.27Issue(7)：1238-1244,7.DOI:10.13313/j.issn.1673-4890.20241204005
基于AS-PCR荧光可视化技术的藏药大籽蒿及其混淆品鉴别研究
Identification of Tibetan Medicine Artemisia sieversiana and Its Adulterant Based on AS-PCR Fluorescence Visualization Technology
摘要
Abstract
Objective:To develop a species-specific DNA fluorescence visualization method for distinguishing Artemisia sieversiana from its adulterant Ajania tenuifolia.Methods:The internal transcribed spacer(ITS),rbcL,and psbA-trnH sequences of Artemisia sieversiana and Ajania tenuifolia were compared.A phylogenetic tree was constructed,and the optimal DNA barcode was selected.Based on the differential sites,specific identification primers for Artemisia sieversiana and Ajania tenuifolia were designed to establish a specific DNA rapid detection system.The amplification system was combined with SYBR Green Ⅰfluorescent dye to achieve fluorescence visualization detection.The established allele-specific polymerase chain reaction(AS-PCR)fluorescence visualization method was used to identify the raw materials in three batches of Wuwei Ganlu Yaoyu Tangsan.Results:ITS was selected as the optimal identification sequence for distinguishing Artemisia sieversiana from Ajania tenuifolia.The AS-PCR fluorescence visualization method based on ITS enabled specific detection within 40 minutes.By using this method to analyze three batches of Wuwei Ganlu Yaoyu Tangsan,Artemisia sieversiana was detected in all three batches,while one batch was found to be adulterated with Ajania tenuifolia.Conclusion:The established method demonstrates good specificity and can be used for the rapid and accurate visualization identification of Artemisia sieversiana and its raw materials in Tibetan medicine preparations.关键词
大籽蒿/细叶亚菊/位点特异性聚合酶链式反应/荧光可视化/藏成药Key words
Artemisia sieversiana Ehrhart ex Willd./Ajania tenuifolia(Jacq.)Tzvel./AS-PCR/fluorescence visualization/Tibetan medicine preparation分类
医药卫生引用本文复制引用
谢雨宸,王亚倩,李慧林,赵程成,陈蓉,范刚..基于AS-PCR荧光可视化技术的藏药大籽蒿及其混淆品鉴别研究[J].中国现代中药,2025,27(7):1238-1244,7.基金项目
国家重点研发计划项目(2023YFC3504403) （2023YFC3504403）
