- Author:
Yu-Qing ZHOU
1
Author Information
- Publication Type:Journal Article
- Keywords: DNA detection kit; DNA extraction; Panax ginseng C. A. Mey; PCR identification; Performance evaluation
- From: Chinese Pharmaceutical Journal 2016;51(21):1866-1870
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To develop a rapid DNA detection kit for DNA extraction and PCR identification of Panax ginseng C. A. Mey. METHODS: The classical DNA extraction and PCR identification methods for Panax ginseng C. A. Mey were modified, and the compositions and reaction conditions of the kit were determined. In addition, the specificity, stability, sensitivity, and repeatability of the kit were evaluated. The genomic DNAs of genuine and counterfeit ginseng goods were extracted by the kit and PCR was performed to identify the authenticity. The purity of the extracted DNA was detected by UV spectrophotometry. Finally, commercially available ginseng samples were verified. RESULTS: The purity of the genomic DNA extracted by the kit was (1.73 ± 0.13) (OD260/OD280), and a fragment between 150 and 200 bp could be amplified only from authentic Panax ginseng C. A. Mey. The specificity of the kit was 100%. The repetitive experiments showed that the average intra-assay CV% and inter-assay CV% of the kit were 2.38% and 2. 62%, respectively. The DNA in solutions diluted by 200 times could still be detected. Stability experiment proved that repeated freeze-thawing for 20 times had no significant effect on the activity of this kit and the test sample could be stored at - 20℃ for one year. The specificity test confirmed that 8 samples among the 10 commercial products were genuine, and 2 were counterfeit. CONCLUSION: The nucleic acid extraction and purity of the DNA detection kit can meet the requirement for identification of Panax ginseng C. A. Mey. The kit has good specificity, high sensitivity, and good stability, so it is suitable for the rapid detection of Panax ginseng C. A. Mey.