中药材哈蟆油PCR鉴定的初步研究

STUDY ON PCR IDENTIFICATION OF THE CHINESE CRUDE DRUG OVIDUCTUS RANAE

从中国林蛙 (Ranachensinensis)、中华大蟾蜍 (Bufogargarizans)以及鳕鱼 (Gadusmacrocephalus)等组织材料中提取DNA ,扩增出约 340bp的 12SrRNA基因片段 ,测定其序列 ,并参考有关序列 ,设计一对哈蟆油的鉴别引物HsmL1、HsmH1,用该鉴别引物扩增蛙类原动物的模板DNA时 ,只有中国林蛙和黑龙江林蛙的模板能得到阳性扩增 ,其余样品为阴性 .因此该对引物能用于哈蟆油药材来源的鉴别 .对购自 5个不同药材商店的哈蟆油PCR鉴定结果表明 ,现市售商品哈蟆油有不同的材料来源 .图 3表 1参

DNA templates were extracted from Rana chensinensis , Bufo gargarizans and Gadus macroccphalu , and 12S rRNA gene fragment was amplified and sequenced with universal primers. Based on the sequence data obtained both from our experiments and the references, a pair of diagnostic primers, HsmL1 and HsmH1, were designed for the authentication of Oviductus Ranae. When PCR was performed using diagnostic primers, positive amplification appeared in the tube with template DNA from R. chensinensis and R. amurensis only. Therefore, HsmL1 and HsmH1 could be used for authentication of Oviductus Ranae. The samples of Oviductus Ranae from 5 different drug stores were examined with the primers, and the results indicate that the sources of material of Oviductus Ranae are still complicated in the markets. Fig 3, Tab 1, Ref 9