| 杨宝,李忠华,董培智,周瑞雪,孙丽萍,裴晓莎,朴晋华.基于高分辨率熔解曲线技术鉴别酸枣仁及其混伪品理枣仁[J].中国药事,2022,36(11):1267-1279 |
| 基于高分辨率熔解曲线技术鉴别酸枣仁及其混伪品理枣仁 |
| Identification of Ziziphi Spinosae Semen and Its Audlterant Ziziphi mauritianae Semen Based on High Resolution Melting Analysis |
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| DOI：10.16153/j.1002-7777.2022.11.009 |
| 中文关键词: 高分辨率熔解曲线 理枣仁 ITS2 酸枣仁 分子鉴定 |
| 英文关键词: high resolution melting Ziziphi mauritianae semen ITS2 Ziziphi spinosae semen molecular identification |
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| 中文摘要: |
| 目的：采用高分辨率熔解曲线技术(HRM)快速鉴别酸枣仁与相关混伪品药材理枣仁及理枣仁杂合品种。方法：采集了14份酸枣仁及10份市售混伪品理枣仁进行样品DNA提取，通过ITS2序列使用NCBI的 Primer-Blast设计引物，筛选引物浓度并对采集的样品进行检测分析。结果：在引物浓度为 10 μmol·L-1的条件下，向反应体系加入0.3μL引物时，可获得优质的熔解曲线。应用该方法对14批酸枣仁及10批理枣仁均可进行HRM曲线分析鉴别，并且通过该方法能够准确区分理枣仁及理枣仁杂合品种；将理枣仁与理枣仁杂合品粉分别掺入酸枣仁样品中，HRM检测技术最低掺伪检测限为10%。结论： 基于HRM技术可实现酸枣仁与其常见混伪品理枣仁、理枣仁杂合品种的准确鉴别，对市场药材的快速检测及质量评估有一定应用前景。 |
| 英文摘要: |
| Objective: To quickly identify Ziziphi Spinosae Semen (ZSS) and its adulterant Ziziphi Mauritianae Semen (ZMS) and the hybrid varieties through high resolution melting curve technique (HRM). Methods: In this study, 14 samples of ZSS and 10 commercial ZMS were collected for DNA extraction. Primers were designed by Primer-Blast of NCBI according to ITS2 sequence to screen the primer concentration and then detect and analyze samples. Results: When the primer concentration was 10 μmol/L, the high quality melting curve could be obtained by adding 0.3 μL primer to the reaction system. A total of 14 samples of ZSS and 10 ZMS could be analyzed and identified through HRM, and the ZMS and its hybrid varieties could be distinguished accurately by this method. When mixing the ZMS and hybrid powder of ZMS in the ZSS samples, the lowest detection limit for adulteration of HRM was 10%. Conclusion: HRM is able to accurately identify ZSS and common adulterant ZMS and its hybrid varieties, which is a promising way in rapid detection and quality evaluation of market medicinal materials. |
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