Jiang Chao, Jin Yan, Zhao Xinlei, Yuan Yuan, Zhao Yuyang, Huang Luqi
a State Key Laboratory Breeding Base of Dao-di Herbs , National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences , Beijing , P.R. China.
b Beijing Medicinal Plant Garden , Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College , Beijing , P.R. China.
Mitochondrial DNA A DNA Mapp Seq Anal. 2018 Mar;29(2):266-272. doi: 10.1080/24701394.2016.1275599. Epub 2017 Jan 10.
Deer antler velvet is widely used as a vitalizing, tonifying, haemopoietic and strengthening agent for debilitated persons in East Asia. To develop a rapid and sensitive method for the identification of the biological source or origin in antler velvet products, a molecular approach was applied using PCR-restriction fragment length polymorphism analysis. The cytochrome b gene sequences of nine cervidae species were analyzed, and a Dde I restriction endonuclease recognition site was found only in sika deer and red deer, the official origin of deer velvet in Chinese pharmacopoeia. A specific primer was designed, and rapid PCR amplified products were subjected to restriction digestion using a fast RFLP procedure. Sika deer and red deer showed two bands of 161 and 102 bp, in contrast to the undigested state of 263 from other antlers. The established PCR-RFLP method was applied in commercial velvet products, and a high frequency of substitution (50%) was revealed in collected commercial samples. The method was successful in detecting contaminated and adulterated antler products in Chinese patent drugs, and the whole detection process was accomplished within 1-1.5 h.
鹿茸在东亚地区被广泛用作滋补强身、补血益气的药物，用于调养虚弱人群。为建立一种快速、灵敏的方法来鉴定鹿茸产品的生物来源或产地，采用了基于PCR-限制性片段长度多态性分析的分子方法。分析了9种鹿科动物的细胞色素b基因序列，发现只有梅花鹿和马鹿（中国药典中鹿茸的法定来源）存在Dde I限制性内切酶识别位点。设计了特异性引物，通过快速RFLP程序对快速PCR扩增产物进行限制性酶切。梅花鹿和马鹿的酶切产物显示出161和102 bp的两条带，而其他鹿茸的未酶切产物为263 bp。将建立的PCR-RFLP方法应用于市售鹿茸产品，结果显示所采集的市售样品中存在较高的替代率（50%）。该方法成功检测出中成药中受污染和掺假的鹿茸产品，整个检测过程可在1-1.5小时内完成。
