利用複合式PCR鑑定倒地蜈蚣藥材及其混淆品   全文下載

Authentication of Dao-Di-U-Gon Medicinal Materials and Its Adulterants Using Multiplex Polymerase Chain Reaction

倒地蜈蚣藥材係瓶爾小草科Ophioglossaceae植物錫蘭七指蕨Helminthostachys zeylanica（L.）Hook.之乾燥根及根莖，是臺灣民間常用中藥。因其具有解熱消炎作用，長期以來被民間傳統醫學廣泛應用。隨著中草藥產品的開發和棲息地的破壞，在許多國家錫蘭七指蕨已被列為瀕危物種，可能導致市場上以其他物種替換或代用的機率提升。另外，由於各地區用藥習慣不同以及名稱相似等原因，倒地蜈蚣藥材之常見混淆品包含地蜈蚣Torenia concolor Lindl.與多羽節肢蕨Arthromeris mairei（Brause）Ching等2個物種。為避免錯誤識別造成之不當使用，建立準確且快速的鑑定方法刻不容緩。近年來分子鑑定方法已被廣泛運用成為識別中草藥品種的有力工具，本研究利用DNA序列差異開發複合式PCR檢測方法，錫蘭七指蕨、地蜈蚣及多羽節肢蕨之特異性擴增產物分別為118、183及93鹼基對（base pair, bp），並經凝膠電泳分離的片段大小來區分倒地蜈蚣藥材及其混淆品，此檢測技術可辨識倒地蜈蚣藥材真偽，並應用於中草藥品質管控，以維護用藥安全。

The root and rhizome of Helminthostachys zeylanica (L.) Hook. (Ophioglossaceae) is a Chinese medicine named“Dao-Di-U-Gon”in Taiwan. It has antipyretic and antiphlogistic effects and has been widely used as folk medicine for a long time. However, with the herbal product development and habitat destruction, H. zeylanica has thus been listed as an endangered species in many countries. Therefore, it may prompt the replacement or substitution with other species in the market. Torenia concolor Lindl. and Arthromeris mairei (Brause) Ching are commonly confusing species due to similar aliases and different medication habits. It is urgent to establish an accurate and rapid identification method to avoid medication errors. In recent years, molecular authentication methods have been widely employed as a powerful tool for the species identification of herbal products. In this study, multiplex PCR was developed for distinguish Dao-Di-U-Gon medicinal materials from its adulterants based on DNA sequence variation. The lengths of the specific amplicons were 118 bp for H. zeylanica, 183 bp for T. concolor, and 93 bp for A. mairei, which could be easily differentiated as three herbal origins by gel electrophoresis. This detection technique can be applied on traditional Chinese medicine (TCM) quality control, thus contributing to medication safety.