中药材2023，Vol.46Issue(9)：2160-2164,5.DOI:10.13863/j.issn1001-4454.2023.09.009
鳖甲配方颗粒的位点特异性PCR鉴别研究
Identification of Trionycis Carapax Formula Granule by Allele-Specific PCR
摘要
Abstract
Objective:To establish a specific polymerase chain reaction(PCR)identification for Trionycis Carapax formula gran-ule.Methods:Based on the cytochrome C oxidase subunit Ⅰ(COXⅠ)sequence of Trionyx sinensis and its adulterants,specific primers BJ-5F,BJ-5R were designed according to the specific single nucleotide polymorphisms(SNP)of Trionyx sinensis.The optimal PCR re-action system was determined by optimizing the PCR reaction procedure,and the tolerability,applicability and sequencing of this method were investigated.Results:When the annealing temperature was 60℃and the cycle number was 33,a single discriminant band at about 236 bp of Trionycis Carapax,standard decoction freeze-dried powder and formula granule could be amplified,while no band was found in other adulterants and negative control.Conclusion:The allele-specific PCR method established in this study can quickly and accurately identify the origin of Trionycis Carapax,standard decoction freeze-dried powder and formula granule,it has the characteristics of strong specificity,high accuracy,simplicity and so on,which provides a reference for the formulation of quality standard of Trionycis Carapax formula granule.关键词
鳖甲/配方颗粒/位点特异性PCR/分子鉴定Key words
Trionycis Carapax/Formula granule/Allele-specific PCR/Molecular identification分类
中医学引用本文复制引用
谭斯尹,宋叶,范耀耀,刘闪闪,胡绮萍,罗宇琴,李国卫..鳖甲配方颗粒的位点特异性PCR鉴别研究[J].中药材,2023,46(9):2160-2164,5.基金项目
广东省基础与应用基础研究基金自然科学基金项目(2020B1515120033) （2020B1515120033）
