利用Nested PCR與DNA定序方法鑑定保育類穿山甲藥材及其製劑

穿山甲藥材是取自保育類動物穿山甲的鱗甲片，為89年11月衛生署公告禁用之藥材，本研究利用PCR與DNA定序方法鑑定中藥製劑是否含有穿山甲成分。將購自藥局之製劑檢體，經過溶解溶液處理，並以另含有SDS的溶解溶液處理穿山甲鱗片檢體，再以有機溶液萃取方法抽取DNA，之後用DNA純化套組的吸附DNA的原理，純化抽取的DNA。再以粒腺體12S rRNA基因作為PCR的模板，利用普遍性引子進行PCR，並設計專一性引子進行nested PCR。nested PCR的產物以自動定序儀定序，將得到的DNA序列與Gen Bank的基因資料庫比對分析，取得鑑定結果。結果顯示，穿山甲藥材山甲片、炮山甲與4件製劑檢體所含的穿山甲成分均為中國穿山甲(Manis pentadactyla)，此為華盛頓公約附錄二所列的保育類動物。

關鍵字

穿山甲； PCR ； nested PCR

並列摘要

A polymerase chain reaction (PCR) with DNA sequencing method has been developed to identify pangolin used in Chinese medicinal raw material and preparations. The samples of pangolin scales, collected from local dispensaries, were treated by lysis buffer, extracted their DNA by organic solution, and then purified using the commercial kit. In this study, we formulated the lysis buffer containing SDS and sarcosyl, and took advantage of the specificity of the kit to purify DNA. Taking mitochondrial 12S rRNA gene as the target template, the modified universal and newly designed primers were used for PCR and for nested PCR respectively. The nested PCR products were sequenced and the obtained sequences were compared with GenBank databases. The results indicated that the species of pangolin in the samples of raw material and preparations were Manis pentadactyla listed in CITES appendix II.

並列關鍵字

pangolin ； PCR ； nested PCR

被引用紀錄

Lu, K. T. (2012). Identification of Botanical Origin in Chinese Medicinal Preparations by Nested PCR and DNA Sequencing Methods. 藥物食品分析, 20(4), 786-793+981. https://doi.org/10.6227/jfda.2012200407

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利用Nested PCR與DNA定序方法鑑定保育類穿山甲藥材及其製劑

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