煅牡蛎(长牡蛎)及其加工产品分子鉴定方法研究  

Molecular authentication of calcined oyster (Ostrea gigas) and its processed products

作　　者：肖智扬 胡力 白云俊 蒋超[1] 赵玉洋 袁媛[2] XIAO Zhi-yang;HU Li;BAI Yun-jun;JIANG Chao;ZHAO Yu-yang;YUAN Yuan(State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室,北京100700 [2]中国中医科学院医学实验中心,北京100700

出　　处：《中国中药杂志》2024年第23期6264-6271,共8页China Journal of Chinese Materia Medica

基　　金：中央级公益性科研院所基本科研业务费专项(ZZ16-ND-12);中国中医科学院科技创新工程项目(CI2023D001,CI2023E002-04);国家杰出青年科学基金项目(82325049);中央本级重大增减支项目(2060302)。

摘　　要：煅牡蛎是我国临床常用贝壳类中药。由于牡蛎在加工过程中,其显微特征遭到破坏,并且明火煅制会破坏牡蛎的DNA,导致难以进行基原鉴定。建立煅牡蛎特异性聚合酶链式反应(PCR)鉴定方法,可为煅牡蛎药材及其加工产品的安全性与临床疗效提供保障。该文以长牡蛎为例,改良了煅牡蛎饮片及其配方颗粒的DNA提取方法,并获得了高质量的DNA;基于长牡蛎与其他2个基原的特异性SNP位点,设计特异性鉴别引物,建立煅牡蛎(长牡蛎)配方颗粒位点特异性鉴别方法,并对该方法进行专属性和适用性考察。结果表明,当退火温度为54℃、循环次数为44次时,煅牡蛎(长牡蛎)及其配方颗粒PCR扩增产物在102 bp处产生单一明亮的鉴别条带,而牡蛎的另外2个来源近江牡蛎、大连湾牡蛎无条带。该研究首次建立了煅法动物类药材DNA提取和PCR鉴定方法,为解决煅法饮片鉴定难的问题、保障用药安全提供工具。Calcined oyster is a commonly used shellfish traditional Chinese medicine in clinical practice in China.During the processing of oysters,their microscopic characteristics are destroyed,and open-fire calcination can damage the DNA of oysters,making it difficult to identify the primary source.The establishment of a specific polymerase chain reaction(PCR)method for the identification of calcined oysters can provide a guarantee for the safety and clinical efficacy of the medicine and its processed products.With Ostrea gigas as an example,the DNA extraction method of decoction pieces and formula particles of calcined oysters was improved,and high-quality DNA was obtained.Based on the specific single nucleotide polymorphism(SNP)sites of O.gigas and the other two species,the specific identification primers were designed,and the site-specific identification method of formula granules of calcined oyster(O.gigas)was established.The specificity and applicability of the method were investigated.The results showed that when the annealing temperature was 54℃,and the cycle was 44 times,the PCR amplified products of calcined oyster(O.gigas)and its formula granules produced a single bright identification band at 102 bp,while the other two species of oysters,O.talienwhanensis Crosse and O.rivularis Gould,had no band.In this study,DNA extraction and PCR identification of animal medicinal materials by calcination were established for the first time,which provided a tool for solving the difficult identification of calcined decoction pieces and ensuring drug safety.

关 键 词：长牡蛎 煅牡蛎 配方颗粒 单核苷酸多态性位点 聚合酶链式反应 

分 类 号：R282.5[医药卫生—中药学]