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1.陕西科技大学 食品科学与工程学院,国家羊乳制品加工技术研发专业中心,陕西 西安 710021
2.山西省实验中学,山西 太原 030032
Received:09 April 2025,
Revised:2025-05-30,
Accepted:01 July 2025,
Published Online:07 November 2025,
Published:15 January 2026
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邓帆,孙娜娜,周凯乐,刘江花,李国梁,徐秦峰,冯霞.基于核酸分子光开关的可视化重组酶聚合酶扩增检测方法[J].分析测试学报,2026,45(01):184-189.
DENG Fan,SUN Na-na,ZHOU Kai-yue,LIU Jiang-hua,LI Guo-liang,XU Qin-feng,FENG Xia.A Visualized Detection Method for Recombinase Polymerase Amplification Based on Nucleic Acid Molecular Light Switch[J].Journal of Instrumental Analysis,2026,45(01):184-189.
邓帆,孙娜娜,周凯乐,刘江花,李国梁,徐秦峰,冯霞.基于核酸分子光开关的可视化重组酶聚合酶扩增检测方法[J].分析测试学报,2026,45(01):184-189. DOI: 10.12452/j.fxcsxb.250409275.
DENG Fan,SUN Na-na,ZHOU Kai-yue,LIU Jiang-hua,LI Guo-liang,XU Qin-feng,FENG Xia.A Visualized Detection Method for Recombinase Polymerase Amplification Based on Nucleic Acid Molecular Light Switch[J].Journal of Instrumental Analysis,2026,45(01):184-189. DOI: 10.12452/j.fxcsxb.250409275.
该研究将核酸分子“光开关”[Ru(bpy)
2
(dppz)]
2+
作为荧光染料应用于实时荧光RPA扩增,实现了对食源性致病菌(如金黄色葡萄球菌(
S.aur
)和沙门氏菌(
S.spp
))的单重和多重实时荧光RPA扩增检测。此外,基于[Ru(bpy)
2
(dppz)]
2+
建立了一种快速、可视化RPA检测方法,该方法通过直接观察扩增产物DNA与[Ru(bpy)
2
(dppz)]
2+
结合后在可见光下产生的强烈红色荧光信号判定扩增结果,避免了高浓度[Ru(bpy)
2
(dppz)]
2+
对扩增反应产生的明显抑制,有良好的可视化检测效果,具有高特异性和高灵敏度,有望广泛应用于环境监测、食品安全和临床诊断的现场快速核酸检测。
Compared with polymerase chain reaction(PCR) and loop-mediated isothermal amplification(LAMP),recombinase polymerase amplification(RPA) has a shorter reaction time(5-20 min) and requires less temperature and equipment,and can be carried out at 37-42 ℃. However,the current organic dyes used for real-time fluorescent RPA are dependent on imports and have small Stoke shifts,which are not conducive to visualization and observation. In this study,we applied the nucleic acid molecule “light switch”[Ru(bpy)
2
(dppz)]
2+
as a fluorescent dye for real-time fluorescent RPA amplification,and realized single and multiple real-time fluorescent RPA amplification detection of food-borne pathogens(e.g.,Staphylococcus aureus(
S.aur
),and Salmonella(
S.spp
)). In addition,a rapid and visualized RPA detection method was established based on[Ru(bpy)
2
(dppz)]
2+
,which determines the amplification results by directly observing the strong red fluorescence signal generated under visible light after the binding of amplification product DNA to[Ru(bpy)
2
(dppz)]
2+
,avoiding the obvious inhibition of the amplification reaction caused by high concentration of[Ru(bpy)
2
(dppz)]
2+
,and has good visualization effect. It avoids the obvious inhibition of the amplification reaction caused by high concentration of [Ru(bpy)
2
(dppz)]
2+
,and has good visualization detection effect,high specificity and high sensitivity,and is expected to be widely used in the field of rapid nucleic acid detection for environmental monitoring,food safety and clinical diagnosis.
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