中国中药杂志

2018, v.43(23) 4562-4568

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多重PCR同时鉴别5种药用海马
Multiplex PCR to simultaneous identification of five traditional Chinese medicinal seahorses

刘富艳;袁媛;金艳;秦雯;蒋超;赵玉洋;黄璐琦;
LIU Fu-yan;YUAN Yuan;JIN Yan;QIN Wen;JIANG Chao;ZHAO Yu-yang;HUANG Lu-qi;State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia, China Academy of Chinese Medical Sciences;Department of Biomedical, Beijing City University;

摘要(Abstract):

海马是我国名贵动物药材之一,《中国药典》收载了5种海马的药用标准。随着海马资源储量的减少,市售海马基原增多,鉴别困难。该研究基于海马及其混伪品线粒体DNA序列差异设计了5对鉴别引物并构建同时鉴别三斑海马、克氏海马、管海马,刺海马和日本海马的多重PCR反应体系。PCR反应结束后进行凝胶电泳,管海马、日本海马、克氏海马、刺海马和三斑海马分别获得155, 222, 292, 352,458 bp的特异性鉴别条带,混伪品无条带。该研究建立的方法可以在单次PCR反应中准确鉴别所有药用海马的物种基原,为海马药材和饮片的质量控制提供了依据。
Seahorse is one the most commonly used medicinal animal in China. Five species of Hippocampus are recorded as seahorse in the Chinese Pharmacopoeia. Because of the rapid decrease, several other Hippocampus species are often adulterants as medicinal seahorse in the herbal market, which compromise clinical efficacy and pose threat to endangered seahorse species conversation. Herein, a multiplex polymerase chain reaction(mPCR) method was developed to identify the biological sources of medicinal seahorses.Based on the sequences of mitochondrial DNA,five specific primers for Hippocampus trimaculatus, H. kelloggi, H. kuda, H. histrix and H. mohnikei(H. japonicus)were designed, respectively. Multiplex PCR yields the products of 155,222,292,352,458 bp amplicons in the present of DNA templates of H. kuda, H. mohnikei, H. kelloggi, H. histrix and H. trimaculatus, respectively. This multiplex PCR method which electrophoresis migration of different lengths of DNA bands allowed simultaneous identification of all the five medicinal seahorses in a single assay. It showed that this multiplex PCR assay is useful for the simultaneous identification the biological sources of complex multi-source samples, which could provide a useful tool for the quality control of seahorses.

关键词(KeyWords): 多重PCR;分子鉴定;海马;中药材;COⅠ序列
multiplex PCR;molecular identification;seahorse;traditional Chinese medicine;COⅠ sequence

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基金项目(Foundation): 中央本级重大增减支项目(2060302);; 中央级公益性科研院所基本科研业务费专项资金项目(ZZ10-008)

作者(Author): 刘富艳;袁媛;金艳;秦雯;蒋超;赵玉洋;黄璐琦;
LIU Fu-yan;YUAN Yuan;JIN Yan;QIN Wen;JIANG Chao;ZHAO Yu-yang;HUANG Lu-qi;State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia, China Academy of Chinese Medical Sciences;Department of Biomedical, Beijing City University;

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