School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Shanghai Insititute for Food and Drug Control, Shanghai, NMPA Laboratory for Quality Control of Traditional Chinese Medicine, Shanghai, China.
Mitochondrial DNA A DNA Mapp Seq Anal. 2020 Jul;31(5):173-177. doi: 10.1080/24701394.2020.1741560. Epub 2020 May 7.
In this study, a method was established for discriminating the true antlers from its counterfeits using TaqMan real-time quantitative PCR. The method combines the use of true antlers-specific primers, that amplify a 226 bp fragment from true antlers DNA, and mammalian-specific primers amplifying a 146 bp fragment from mammalian species DNA, which are used as endogenous control. A TaqMan probe that hybridizes in the ' antler' and also in the 'mammalian' DNA fragments is used to monitor the amplification of the target gene. The antler mitochondrial DNA was used as target gene to design the primers and TaqMan probes. The data revealed that the TaqMan real-time PCR-based assay can be used for identification of the true antlers from counterfeits in a single step. The limit of detection (LOD) was lower than 1 pg of DNA per reaction.
在这项研究中，建立了一种使用 TaqMan 实时定量 PCR 鉴别真假鹿茸的方法。该方法结合使用真鹿茸特异性引物，从真鹿茸 DNA 中扩增 226bp 片段，以及哺乳动物特异性引物，从哺乳动物 DNA 中扩增 146bp 片段，作为内参。使用 TaqMan 探针杂交在“鹿茸”和“哺乳动物”DNA 片段中，以监测靶基因的扩增。选择鹿线粒体 DNA 作为靶基因来设计引物和 TaqMan 探针。结果表明，该 TaqMan 实时 PCR 检测法可用于一步鉴别真假鹿茸。检测限（LOD）低于每个反应 1pg 的 DNA。
