Development and evaluation of a rapid PCR detection kit for Ophiocordyceps sinensis.
10.19540/j.cnki.cjcmm.20170217.011
- Author:
Fei-Xia HOU
1
;
Jing CAO
1
;
Sha-Sha WANG
1
;
Xi WANG
1
;
Yuan YUAN
2
;
Cheng PENG
1
;
De-Guang WAN
1
;
Jin-Lin GUO
1
Author Information
1. Pharmacy College, Chengdu University of Traditional Chinese Medicine, the Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine, Key Laboratory of Systematic Research, Development and Utilization of Chinese Medicine Resources in Sichuan Province-Key Laboratory Breeding Base of Co-founded by Sichuan Province and MOST, Chengdu 611137, China.
2. State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Ophiocordyceps sinensis;
PCR;
detection kit
- From:
China Journal of Chinese Materia Medica
2017;42(6):1125-1129
- CountryChina
- Language:Chinese
-
Abstract:
Ophiocordyceps sinensis is a valuable traditional Chinese medicine. Due to resource shortage, expensive price and huge market demand, there are many adulterants of O. sinensis in markets. Therefore, it is necessary to establish a rapid and effective method for distinguishing O. sinensis. Based on the species-specific PCR of O. sinensis, this study developed a detection kit by optimizing the components and evaluated the specificity, detection limit, repeatability and shelf life of the kit. The results showed that when the quality of O. sinensis accounted for more than 1/200 of that mixture, it could be detected successfully. Moreover, only O. sinensis could be amplified and glowed bright green fluorescence under ultraviolet light. The kit was still in effect when it was placed at 37 ℃ for three days, which indicated that it was stable and effective for one year stored in 4 ℃. The kit in the same batch under different operation conditions, and in different batch under the same operation conditions gave the same result and accuracy, which showed good repeatability of the kit. It is simple, rapid and accurate to distinguish O. sinensis from its adulterants using the kit, and lays the foundation for commercialization of traditional Chinese medicine fast detection kit.
