摘要
目的:利用位点特异性聚合酶链式反应(PCR),建立一种能够快速鉴别当归药材及饮片中掺伪欧当归的方法。方法:通过比对当归和欧当归ITS基因序列,寻找单核苷酸多态性(SNP)位点并设计特异性鉴别引物。对影响特异性PCR反应的退火温度、引物浓度、循环数等条件进行优化,对掺伪不同比例和不同来源的当归掺伪样品进行了专属性、准确性和稳定性考察。结果:所建立的位点特异性PCR鉴别方法,在退火温度为63℃、引物循环数为30时,掺有欧当归的当归样品经过特异性引物扩增后,在250~500 bp之间检出一条单一DNA条带,而当归样品则无此条带。结论:该研究建立的位点特异性PCR方法能够准确地检测出当归药材及饮片中是否掺有欧当归,可作为当归质量监测的新手段。
Objective:To establish a rapid method to identify Angelica sinensis and its adulteration Levisticum officinale by site-specific polymerase chain reaction(PCR).Methods:By comparing the ITS gene sequences of Angelica sinensis and Levisticum officinale,the single nucleotide polymorphism(SNP)sites were searched and specific identification primers were designed.The annealing temperature,primer concentration,cycle number and other conditions affecting the specific PCR reaction were optimized.The specificity,accuracy and stability of Angelica sinensis mixed with Levisticum officinale with different proportion and different sources were investigated.Results:A site-specific PCR method for identifying Angelica sinensis mixed with Levisticum officinale was established.When the annealing temperature was 63℃and the number of primer cycles was 30,a single DNA band of 250~500 bp was detected in the Angelica sinensis mixed with Levisticum officinale after amplification with specific primers,while Angelica sinensis was negative.Conclusion:The established site-specific PCR method can accurately detect whether there is Levisticum officinale in Angelica sinensis,which can be used as a new method for quality monitoring of Angelica sinensis.
作者
史中飞
滕宝霞
倪琳
赖晶
宋平顺
SHI Zhong-fei;TENG Bao-xia;NI Lin;LAI Jing;SONG Ping-shun(Gansu Institute for Drug Control/National Key Laboratory of Quality Control of Chinese Medicinal Materials and Decoction Pieces,Lanzhou 730070,China)
出处
《中药材》
CAS
北大核心
2021年第7期1597-1602,共6页
Journal of Chinese Medicinal Materials
基金
甘肃省药品监督管理局药品科研项目(2020GSMPA024)
国家重点研发计划“中医药现代化研究”重点专项(2019YFC1711500)
关键词
当归
欧当归
特异性PCR
掺伪鉴别
Angelica sinensis(Oliv.)Diels
Levisticum officinale Koch
Specific PCR
Adulterated identification
