| 基于LNA-TaqMan探针实时荧光定量PCR检测技术的天山雪莲掺伪研究 |
| 投稿时间：2024-09-13 修订日期：2024-11-18 点此下载全文
|
| 引用本文： |
| DOI： |
| 摘要点击次数: 54 |
| 全文下载次数: 0 |
|
|
| 基金项目:安徽省药品监督管理局科技创新项目 |
|
| 中文摘要:目的：采用LNA-TaqMan实时荧光定量聚合酶链式反应（PCR）技术建立一种高效、快速鉴定天山雪莲中掺伪水母雪兔子、绵头雪兔子的方法。方法：基于锁核酸（LNA）-TaqMan探针实时荧光定量PCR技术，利用天山雪莲及其常见易混伪品水母雪兔子、绵头雪兔子的叶绿体基因组序列差异，根据伪品水母雪兔子、绵头雪兔子与天山雪莲共有的特异性差异位点设计筛选探针引物，并对引物及LNA-TaqMan探针的特异性进行验证，根据扩增曲线Ct值的差值，计算掺伪比例。结果：基于LNA-TaqMan探针检测方法，通过出现特异性扩增曲线判定检测出水母雪兔子和绵头雪兔子，根据扩增曲线Ct值的差值能有效确定掺伪比例，且在掺伪1%时，仍可稳定检出。结论：该方法用于水母雪兔子和绵头雪兔子掺伪天山雪莲检测灵敏度高，特异性强，可有效解决天山雪莲混伪品难以鉴别的问题，为天山雪莲药材质量控制提供技术支持。 |
| 中文关键词:天山雪莲 水母雪兔子 绵头雪兔子 锁核酸-TaqMan探针 实时荧光定量PCR 掺伪 |
| |
| Adulteration detection of saussurea involucrata Based on the real-time fluorescence quantitative PCR technology of LNA-TaqMan probe |
|
|
| Abstract:Objective: A highly efficient and rapid method for identifying Saussurea?medusa and Saussurea?laniceps in saussurea involucrata was established using LNA TaqMan real-time fluorescence quantitative polymerase chain reaction (PCR) technology.Methods: Based on the real-time fluorescence quantitative PCR technology of Lock Nucleic Acid (LNA) TaqMan probe, the chloroplast genome sequence differences of Saussurea involucrata and its common easily mixed Saussurea?medusa and Saussurea?laniceps were utilized to design and screen probe primers according to the specific differences shared by Saussurea?medusa，Saussurea?laniceps and Saussurea involucrata. The specificity of primers and LNA TaqMan probe was verified, and the doping ratio was calculated based on the difference in Ct values of amplification curves.Results:Based on the LNA TaqMan probe detection method, the detection of Saussurea?medusa and Saussurea?laniceps can be determined by the appearance of specific amplification curves. The doping ratio can be effectively determined by the difference in Ct values of the amplification curves, and stable detection can still be achieved at 1% doping. Conclusion:This method has high sensitivity and specificity for detecting counterfeit Saussurea involucrata in Saussurea?medusa and Saussurea?laniceps, and can effectively solve the problem of difficult identification of Saussurea involucrata mixed products, providing technical support for the quality control of Saussurea involucrata medicinal materials. |
| keywords:Saussurea involucrata Saussurea?medusa? Saussurea?laniceps LNA-Taqman probe real-time fluorescent quantitative PCR adulteration |
|
查看全文
查看/发表评论 下载PDF阅读器 |
