Authentication of animal signatures in traditional Chinese medicine of Lingyang Qingfei Wan using routine molecular diagnostic assays
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- 作者：Meng Cao (1)
Jikun Wang (1)
Lu Yao (2)
Suhua Xie (2)
Jing Du (2)
Xingbo Zhao (1) - 关键词：Lingyang Qingfei Wan ; Saiga tatarica ; Capra hircus ; Molecular diagnostic assay ; Sample preprocessing
- 刊名：Molecular Biology Reports
- 出版年：2014
- 出版时间：April 2014
- 年：2014
- 卷：41
- 期：4
- 页码：2485-2491
- 全文大小：1,100 KB
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Jikun Wang (1)
Lu Yao (2)
Suhua Xie (2)
Jing Du (2)
Xingbo Zhao (1)
1. National Engineering Laboratory for Animal Breeding, Ministry of Agricultural Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, China
2. Research Institute of Beijing Tongrentang Co., Ltd, Beijing, 100079, China - ISSN：1573-4978
文摘
Lingyang Qingfei Wan produced by Beijing TongRenTang is a long-standing and popular medicine in China and international pharmaceutical markets. Concerns continue to be raised about the legality of usage of saiga antelope, which was defined as endangered species by Convention on International Trade in Endangered Species of Wild Fauna and Flora legislation and internal legislation in China. Therefore, the alternative pill in which substitutes saiga antelope with goat in the formula of Lingyang Qingfei Wan was developed. In order to authenticate the origin of animal contents in Lingyang Qingfei Wan and its alternative pill, molecular diagnostic assay was utilized by mtDNA polymorphism analysis. Four universal primer pairs containing mtDNA 12SrRNA, 16SrRNA, cytochrome b gene and cytochrome oxidase I were employed to obtain species-specific sequences of saiga antelope and goat, and multiple species-specific primer pairs for saiga antelope and goat were used to identify the animal origin in patent pills according to nucleotide polymorphisms between the two species. In additions, alternative techniques were attempted surrounding dilemmas of low concentration of target DNAs and presence of PCR-inhibitory substances in organic ingredients within complex pill. Results revealed that all species-specific primers could be successfully used for authentication of animal origin within complex pill, and sample preprocessing was critical during experimental manipulation. Internal positive control was an efficient and cost-effective way to assist in monitoring the potential interference from inhibitory substances which existed in the highly processed pills.