中国现代中药2024，Vol.26Issue(3)：457-462,6.DOI:10.13313/j.issn.1673-4890.20230829001
基于LNA-TaqMan探针实时荧光定量PCR检测技术的药用黄精掺伪研究
Adulteration Detection of Polygonati Rhizoma Based on Real-time Fluorescence Quantitative PCR Technology with LNA-TaqMan Probe
摘要
Abstract
Objective:A rapid,sensitive,and efficient real-time polymerase chain reaction(PCR)approach was developed in this work in order to detect the adulteration of Polygonatum zanlanscianense Pamp with Polygonati Rhizoma.Methods:Based on Locked Nucleic Acid(LNA)-TaqMan probe real-time fluorescence quantitative PCR technology,the sequence differences of the trnC-petN gene of chloroplast DNA from various original samples of Polygonati Rhizoma was employed in this study to designe and screen specific primers and probes of specific differential sites for common adulterants.The specificity of the primers and LNA-TaqMan probes(Locked nucleic acid probes)were validated.The adulteration ratio of P.zanlanscianense with Polygonati Rhizoma was calculated according to the difference in Ct values of the amplification curve.Results:The results indicated that the real-time fluorescence quantitative PCR technology with LNA-TaqMan probe detection method can specifically detect the adulteration of P.zanlanscianense with Polygonati Rhizoma and determine the adulteration ratio.Stable detection was achieved even at 1%adulteration level.Conclusion:The method is simple,accurate,reproducible and stable,and can be used for the quantitative detection of Polygonati Rhizoma adulterated with P.zanlanscianense.关键词
湖北黄精/锁核酸-TaqMan探针/实时荧光定量聚合酶链式反应/掺伪/鉴定Key words
Polygonatum zanlanscianense Pamp/LNA-Taqman probe/real-time fluorescent quantitative PCR/adulteration/identification分类
医药卫生引用本文复制引用
王多梅,胡冲,蒲婧哲,陈灵丽,杨建波,张亚中,张文娟..基于LNA-TaqMan探针实时荧光定量PCR检测技术的药用黄精掺伪研究[J].中国现代中药,2024,26(3):457-462,6.基金项目
安徽省药品监督管理局监管科学研究重点项目(AHYJ-KJ-202209) （AHYJ-KJ-202209）
常见与重要中药材及饮片DNA分子鉴定研究项目(TCM2021-YJ07) （TCM2021-YJ07）
