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Identification of Ophiocordyceps sinensis and its adulterants based on portable and CFX96 real-time fluorescent PCR systems
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Kai-yi YUN1, 2, Li XIANG3, Xiao-yue WANG1, 2, Yang LIU1, 2, Hui YAO1, 2, Jing-yuan SONG1, 2, *
Acta Pharmaceutica Sinica | 2019, 54(4) : 746 - 752
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Acta Pharmaceutica Sinica | 2019, 54(4): 746-752
• Original Articles •
Identification of Ophiocordyceps sinensis and its adulterants based on portable and CFX96 real-time fluorescent PCR systems
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Kai-yi YUN1, 2, Li XIANG3, Xiao-yue WANG1, 2, Yang LIU1, 2, Hui YAO1, 2, Jing-yuan SONG1, 2, *
Affiliations
  • 1. Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
  • 2. Engineering Research Center of Chinese Medicine Resources, Ministry of Education, Beijing 100193, China
  • 3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
Published: 2019-04-12 doi: 10.16438/j.0513-4870.2019-0078
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The molecular identification of Ophiocordyceps sinensis and its adulterants was carried out by real-time fluorescent PCR with TaqMan probe. Genomic DNA was extracted from 100 samples of Ophiocordyceps sinensis and its adulterants. MEGA 7.0 software was used for comparative analysis to define the variable sites between Ophiocordyceps sinensis and its adulterants according to the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). A set of specific primers and TaqMan probe were designed using Primer Premier 6.0 software, and sensitivity and specificity studies were performed on two different real-time fluorescent PCR systems (Genesig q16 and Bio-Rad CFX96). The sensitivity study showed that the detectable DNA template concentration of Ophiocordyceps sinensis for the real-time fluorescent PCR was 0.016 ng·μL-1 in the Bio-Rad CFX96 system and 15.527 ng·μL-1 in the Genesig q16 system, respectively. Meanwhile, this method had good specificity for Ophiocordyceps sinensis on Genesig q16 and Bio-Rad CFX96 systems, so Ophiocordyceps sinensis could be clearly distinguished from Ophiocordyceps nutans, Cordyceps gunnii, Cordyceps militaris, Cordyceps cicadae, Cordyceps liangshanensis, Cordyceps gracilis. Our results indicate that real-time fluorescent PCR with TaqMan probe can be used to accurately identify Ophiocordyceps sinensis from its adulterants. This provides a technical method that has wide applications for market management and quality control of Chinese materia medica.

Ophiocordyceps sinensis  /  TaqMan probe  /  real-time fluorescent PCR  /  molecular identification
Kai-yi YUN, Li XIANG, Xiao-yue WANG, Yang LIU, Hui YAO, Jing-yuan SONG. Identification of Ophiocordyceps sinensis and its adulterants based on portable and CFX96 real-time fluorescent PCR systems[J]. Acta Pharmaceutica Sinica, 2019 , 54 (4) : 746 -752 . DOI: 10.16438/j.0513-4870.2019-0078
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Article Info
doi: 10.16438/j.0513-4870.2019-0078
  • Receive Date：2019-01-28
  • Online Date：2026-01-26
  • Published：2019-04-12
Article Data
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History
  • Received：2019-01-28
  • Revised：2019-02-22
Funding
Affiliations
    1. Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
    2. Engineering Research Center of Chinese Medicine Resources, Ministry of Education, Beijing 100193, China
    3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
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表12种不同金属材料的力学参数
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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