醋龟甲配方颗粒的位点特异性PCR鉴别  

Identification of Vinegar Testudinis Carapax et Plastrum Formula Granules by Allele-Specific PCR

作　　者：谭斯尹 潘礼业 宋叶 刘闪闪 范耀耀 罗宇琴 李国卫 TAN Si-yin;PAN Li-ye;SONG Ye;LIU Shan-shan;FAN Yao-yao;LUO Yu-qin;LI Guo-wei(Guangdong Yifang Pharmaceutical Co.,Ltd.,Foshan 528244,China;Guangdong Key Laboratory of TCM Formula Granule Enterprises,Foshan 528244,China)

机构地区：[1]广东一方制药有限公司,广东佛山528244 [2]广东省中药配方颗粒企业重点实验室,广东佛山528244

出　　处：《中国现代中药》2024年第8期1392-1398,共7页Modern Chinese Medicine

基　　金：工业和信息化部2022年产业技术基础公共服务平台项目(2022-230-221);2022年佛山市南海区重点领域科技攻关专项(南科〔2023〕20号-18)。

摘　　要：目的:建立一种醋龟甲配方颗粒特异性位点聚合酶链式反应(PCR)鉴别方法。方法:根据乌龟Chinemys reevesii(Gray)及其混伪品的细胞色素C氧化酶亚基Ⅰ基因序列上的单核苷酸多态性位点设计醋龟甲的特异性鉴别引物,经过优化反应体系,确定最佳的PCR反应条件,并对该方法进行耐受性和适用性考察。结果:当退火温度为62℃、循环数为34时,醋龟甲饮片、标准汤剂冻干粉和配方颗粒均可在约92 bp处得到清晰单一的特异性条带,其混伪品及阴性对照均无条带。结论:建立的位点特异性PCR鉴别方法可快速、准确地鉴别醋龟甲饮片、标准汤剂冻干粉和配方颗粒的真伪,为醋龟甲配方颗粒质量标准研究提供了参考。Objective:To establish an allele-specific polymerase chain reaction(PCR)identification method for vinegar Testudinis Carapax et Plastrum formula granules.Methods:Based on the single nucleotide polymorphism on the cytochrome C oxidase subunitⅠgene sequence of Chinemys reevesii(Gray)and its adulterant,specific primers of vinegar Testudinis Carapax et Plastrum were designed.The optimal PCR reaction system was determined by optimizing the reaction system,and the tolerability and applicability of this method were investigated.Results:When the annealing temperature was 62℃,and the cycle number was 34,the specific band of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules could be obtained at about 92 bp,while no band was found in other adulterants and negative control.Conclusion:The established allele-specific PCR identification method can quickly and accurately identify the authenticity of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules,providing a reference for the research on quality standards of vinegar Testudinis Carapax et Plastrum.

关 键 词：醋龟甲 配方颗粒 位点特异性聚合酶链式反应 分子鉴定 

分 类 号：R282.12[医药卫生—中药学]