基于荧光共振能量转移的核酸适配体传感器检测环丙沙星  

A fluorescence resonance energy transfer-based aptasensor for the detection of ciprofloxacin

作　　者：李滨汐 王婷 王安琪 王雪 钱美汝 朱咏莉 李胎花 LI Binxi;WANG Ting;WANG Anqi;WANG Xue;QIAN Meiru;ZHU Yongli;LI Taihua(CoInnovation Center for the Sustainable Forestry in Southern China,College of Biology and the Environment,Nanjing Forestry University,Nanjing 210037,China;National Positioning Observation Station of Hung-tse Lake Wetland Ecosystem in Jiangsu Province,Hongze 223100,China)

机构地区：[1]南京林业大学生物与环境学院南方现代林业协同创新中心,南京210037 [2]江苏洪泽湖湿地生态系统国家定位观测研究站,洪泽223100

出　　处：《分析试验室》2023年第11期1483-1488,共6页Chinese Journal of Analysis Laboratory

基　　金：江苏省自然科学基金(BK20191387)项目资助。

摘　　要：建立了一种基于荧光共振能量转移(FRET)的核酸适配体传感器,并用于检测实际水体和牛奶中的环丙沙星(CIP)。为了防止羧基荧光素(FAM)被CIP猝灭,FAM和四甲基罗丹明(TAMRA)分别标记在互补单链DNA(FAM-cDNA)和适配体(TAMRA-APT),通过DNA杂交发生FRET,TAMRA有效猝灭FAM的荧光。CIP加入后,其与FAM-cDNA发生亲和力竞争反应,CIP与TAMRA-APT形成结构更稳定的CIP/TAMRA-APT复合物,使体系FAM的荧光恢复。在优化条件下,本方法对CIP表现出高灵敏度和高选择性,检测浓度线性范围为0.01~1μmol/L,检出限为6 nmol/L;对实际水样和牛奶的加标回收率为90.4%~113.2%,相对标准偏差为1.8%~11%。该荧光适配体传感器具有成本低、灵敏度高、特异性好等优点,在环境中CIP残留快速检测方面具有良好的应用潜力。A fluorescence resonance energy transfer(FRET)-based aptasensor was established for the detection of ciprofloxacin(CIP)in water and milk.The fluorescence of carboxyfluorescein(FAM)could be effectively quenched by tetramethylrhodamine(TAMRA)based on the FRET occurred by the hybridization of TAMRA-labeled aptamer(TAMRA-APT)with fluorescein-labeled complementary DNA(FAM-c DNA).However,CIP could compete with FAM-c DNA in affinity,which means after its addition to form a more stable CIP/TAMRA-APT complex with TAMRA-APT,FAM-c DNA was released,leading to the fluorescence of FAM system recovered.As a result,the aptasensor showed high sensitivity and selectivity for CIP detection.Under the optimal conditions,the linear range of CIP concentrations for the method was 0.01-1μmol/L,and the detection limit was 6 nmol/L.Furthermore,the recoveries of the spiked water and milk samples were 90.4%-113.2%,and the relative standard deviations(RSDs)were 1.8%-11%.The fluorescent aptasensor,with low cost,high sensitivity and specificity,has good application potential in the rapid detection of CIP residues in the environment.

关 键 词：适配体传感器 环丙沙星 氟喹诺酮类抗生素 荧光共振能量转移 荧光猝灭 

分 类 号：X52[环境科学与工程—环境工程] X56