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OBJECTIVE To establish a molecular identification method for the adulteration of Zaocys dhumnades in Chinese patent medicine. METHODS Eleven kinds of Chinese patent medicines containing Zaocys dhumnades and 15 kinds of Zaocys dhumnades and their adulterants were collected. The total genomic DNA was extracted from all samples, and the comparison and analysis of Zaocys dhumnades and its adulterants were conducted based on 12S gene fragments. The universal primer sequence, the specific primer sequence and the primers and probes used for fluorescence quantification of Zaocys dhumnades were designed according to the difference points. Different PCR methods were used to amplify the DNA of Zaocys dhumnades contained in the medicinal materials of Zaocys dhumnades and Chinese patent medicines respectively. The reaction system was optimized and the tolerance and adaptability of the methods were investigated. RESULTS The specific PCR technique of Zaocys dhumnades was established. All the authentic Zaocys dhumnades had a clear band around 200 bp, and other adulterants had no band. Fluorescent quantitative PCR method was established for the identification of Chinese patent medicines with low DNA content. CONCLUSION The medicinal materials of Zaocys dhumnades can be effectively identified by PCR amplification with specific primers. Fluorescent quantitative PCR detection can effectively identify the authenticity of Zaocys dhumnades in Chinese traditional medicine.The method is specific, rapid and sensitive, which provides a reference for the identification of Zaocys dhumnades in Chinese patent medicine.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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