建立一种用于快速鉴定冬凌草及其混淆品的种属特异性多重 PCR 和 DNA 测序方法。
Development of the species-specific multiplex PCR and DNA sequencing methods for rapid authentication of Isatidis Folium and its adulterants.
机构信息
Division of Research and Analysis, Food and Drug Administration, Ministry of Health and Welfare, No.161-2 Kunyang St., Nangang Dist., Taipei 11561, Taiwan.
Southern Center for Regional Administration, Food and Drug Administration, Ministry of Health and Welfare, No.180, Zihyou 2nd Rd., Zuoying Dist., Kaohsiung 81358, Taiwan.
出版信息
J Food Drug Anal. 2021 Jun 15;29(2):303-310. doi: 10.38212/2224-6614.3354.
Isatis indigotica Fort. (family Cruciferae), is an herb widely used in traditional herbal medicine and its dried leave was named "ISATIDIS FOLIUM". Baphicacanthus cusia (Ness) Bremek. and Polygonum tinctorium Ait. are commonly misused as ISATIDIS FOLIUM in Chinese Medicine pharmacy. For the purpose of being not misused, specific primers based on the sequence difference of chloroplast trnH-psbA intergenic spacer were designed and multiplex polymerase chain reaction method (multiplex PCR) was developed. In this study, 29 original herbal materials were analyzed and our results show that DNA size after multiplex PCR was able to distinguish variations between three herbs. DNA fragments of 464, 297, 170 base pairs (bps) were represented for I. indigotica and B. cusia and P. tinctorium, respectively. In conclusion, our investigations demonstrate that molecular identification method provides more accurate results for medicinal plants detection and good quality control of ISATIDIS FOLIUM.
菘蓝（十字花科）是一种广泛应用于传统草药的草本植物，其干燥的叶子被称为“菘蓝叶”。白花蛇舌草（Ness）Bremek. 和蓼科植物蓼蓝（Polygonum tinctorium Ait.）常被误用为中药店的菘蓝叶。为了防止误用，根据叶绿体 trnH-psbA 基因间隔区序列差异设计了特定引物，并开发了多重聚合酶链反应（multiplex PCR）方法。在这项研究中，分析了 29 种原始草药材料，结果表明，多重 PCR 后的 DNA 大小能够区分这三种草药的变异。464、297、170 碱基对（bps）的 DNA 片段分别代表菘蓝、白花蛇舌草和蓼蓝。总之，我们的研究表明，分子鉴定方法为药用植物的检测提供了更准确的结果，并对菘蓝叶的质量控制具有良好的效果。
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