人参实时荧光定量PCR鉴定方法的建立

Development of Real-Time PCR for Identification of Panax ginseng

根据人参和人参属其他物种的18S rRNA基因序列差异设计特异引物和探针,建立人参TaqMan实时荧光PCR鉴定方法.应用该方法测试58个样品,包括30个不同来源的人参样品、 15个其他人参属样品以及13个形态近似样品.结果表明:所有供试的人参样品均表现为阳性扩增,非人参及空白对照均未出现阳性扩增.灵敏度检测结果显示:此实时荧光PCR方法的最低检测限为0.03 pg/μL反应.此方法可快速、准确、灵敏地鉴定人参及其加工产品,适用于口岸进出口鉴定.

In this paper, a TaqMan real-time PCR method for identification of Panax ginseng was developed with specific primers and probe designed from the 18S ribosomal RNA gene of P. ginseng. A total of 58 samples including 30 P. ginseng samples of various sources, 15 other Panax sp. samples and 13 samples having similar appearance with P. ginseng were tested with the probe and primers for Taq Manreal-time PCR. Results showed that typical amplification curves could be obtained from all the 30 P. ginseng samples, but not from other 28 samples and blankcontrol. Sensitivity of this method reached to 0.03 pg/μL. The established TaqMan real-time PCR technique in this study enables rapid, accurate and sensitive identification of P. ginseng, which is suitable for the application for customs surveillance.