•  
  •  
  •  
  •  
  •  
  •  
  •  
  •  
  •  
期刊检索


检索
检索项:
检索词:
总目录
  • WCSB9︱2019(第九届)世界采样和混样大会 查看全文>>
  • 关于召开第十五届全国青年分析测试学术报告会的通知 查看全文>>
友情链接
基于核酸分子光开关的闭管可视化环介导等温扩增检测方法
    点此下载全文
作者单位
王芳,董菁,李艳妮,徐秦峰 陕西科技大学 食品与生物工程学院陕西 西安 710021 
基金项目:国家自然科学基金资助项目(22074085);国家重点研发计划子课题(2019YFC1606302)
中文摘要:该研究采用核酸分子“光开关”[Ru(bpy)2(dppz)]2+作为环介导等温扩增(LAMP)荧光染料,建立了一种快速、直观的闭管可视化LAMP检测方法。为避免高浓度染料对LAMP反应造成的强烈抑制和开盖导致的气溶胶交叉污染,该文采用蜡封反应管进行检测,通过直接观察扩增产物DNA与[Ru(bpy)2(dppz)]2+结合后产生的强烈红色荧光信号判定结果。该方法对金黄色葡萄球菌DNA的检出限可低至20拷贝/反应。方法特异性强,整个可视化检测过程可在1 h内完成,有望广泛应用于环境检测、食品安全、临床诊断等领域的现场快速核酸检测。
中文关键词:环介导等温扩增法(LAMP)  ?[Ru(bpy)2(dppz)]2+  ?可视化检测  ?金黄色葡萄球菌
 
A Visualized Closed-tube Detection Method for Loop-mediated Isothermal Amplification Based on Nucleic Acid Molecular Light Switch
Abstract:Compared with polymerase chain reaction(PCR) nucleic acid amplification,loop-mediated isothermal amplification(LAMP) does not require thermal cycling instruments,which is more suitable for rapid on-site detection.However,the small Stoke′s shift of currently used organic fluorescent dye for visualized LAMP leads to insufficient color differentiation.In this study,the nucleic acid molecule“light switch”[Ru(bpy)2(dppz)]2+ was exploited as a LAMP fluorescent dye to establish a rapid visualized closed-tube method for LAMP detection.Visualization detection requires a high concentration of dye,but its adding before amplification reaction will strongly inhibit LAMP reaction,while its adding after reaction by opening the lid will lead to a serious aerosol cross contamination.Therefore,wax was chosen to seal the[Ru(bpy)2(dppz)]2+ at the bottom of the tube,and the LAMP amplification products were visually detected by using the strong red fluorescence emission of [Ru(bpy)2(dppz)]2+ in the presence of double-strand DNA.The S.aureus DNA could be detected as low as 20 copies/reaction with strong specificity,and the whole visual detection process could be completed within 1 h.It could be widely used for the rapid on-site nucleic acid detection in the fields of environmental detection,food safety and clinical diagnosis.
Key Words:LAMP isothermal amplification  ?[Ru(bpy)2(dppz)]2+  ?visual detection  ?S. aureus
引用本文:王芳,董菁,李艳妮,徐秦峰.基于核酸分子光开关的闭管可视化环介导等温扩增检测方法[J].分析测试学报,2022,41(4):646-651.
摘要点击次数: 94
全文下载次数: 199
查看全文  下载PDF阅读器