基于发光共振能量转移的无标记上转换荧光探针灵敏检测8-OHdG  

Sensitive Detection of 8-OHdG by a Label-free Upconversion Fluorescent Probe Based on Luminescence Resonance Energy Transfer

作　　者：闾心悦 于慧 曹丹 郭会琴[1] 颜流水[1] 刘小明[1] LYU Xin-yue;YU Hui;CAO Dan;GUO Hui-qin;YAN Liu-shui;LIU Xiao-ming(School of Environmental and Chemical Engineering,Nanchang Hangkong University,Nanchang 330063,China;Jinhua Customs Comprehensive Technology Service Center,Zhejiang Jinhua 321001,China)

机构地区：[1]南昌航空大学环境与化学工程学院,南昌330063 [2]金华海关综合技术服务中心,浙江金华321001

出　　处：《南昌航空大学学报（自然科学版）》2023年第3期76-87,共12页Journal of Nanchang Hangkong University(Natural Sciences)

基　　金：国家自然科学基金(21665018,21966023);江西省自然科学基金(20181BAB203014)。

摘　　要：8-羟基-2’-脱氧鸟苷(8-hydroxy-2-deoxyguanosine,8-OHdG)是一种重要的DNA损伤标志物,其含量的多少可反映人体的氧化损伤程度。本研究以氨基功能化的上转换纳米粒子(Upconversion nanoparticles,UCNPs)为能量供体,金纳米粒子(AuNPs)为能量受体,由于AuNPs可导致UCNPs在545 nm处的荧光发生猝灭,而8-OHdG能够对AuNPs产非特异性诱导团聚作用,从而改变UCNPs在该位置处的荧光强度,据此构建了一种基于发光共振能量转移(Luminescence resonance energy transfer,LRET)的上转换荧光探针,并用于尿样基质中8-OHdG的检测。所建立方法简单、灵敏且适用pH范围宽。在AuNPs溶液的浓度为1.34 nmol•L^(−1),pH为2~7的实验条件下,所构建探针能够分别在低浓度范围(1.67×10^(−11)~3.3×10^(−9)mol•L^(−1))和高浓度范围(1.0×10^(−8)~3.3×10^(−6)mol•L^(−1))内实现对8-OHdG的快速检测。共存干扰实验结果表明,尿样中常见的无机离子和化合物存在的前提下,所构建探针对8-OHdG呈现了良好的选择性。将探针应用于实际尿液样品基质中8-OHdG的检测,加标回收率为91.4%~109.2%。8-Hydroxy-2’-deoxyguanosine(8-OHdG)is an important DNA damage marker,which can reflect the degree of oxidative damage in human body.In this study,a LRET-based label-free upconversion fluorescent probe was constructed based on the Luminescence resonance energy transfer between amino-functionalized upconversion nanoparticles(UCNPs)as the energy donor and gold nanoparticles(AuNPs)as the energy acceptor.As the AuNPs can induce the fluorescence quenching of UCNPs at 545 nm,and 8-OHdG can produce nonspecifically induced clustering of AuNPs,the fluorescence intensity of UCNPs is altered,which renders the detection of 8-OHdG in urine matrix.The developed method is simple,sensitive,and workable over a wide pH window.When the concentration of AuNPs solution is 1.34 nmol•L^(−1)and the pH is 2~7,the 8-OHdG can be sensitively detected both in the low concentration range(1.67×10^(−11)~3.3×10^(−9)mol•L^(−1))and high concentration range(1.0×10^(−8)~3.3×10^(−6)mol•L^(−1)).The anti-interference experiment results show that the co-existing inorganic ions and organic compounds found commonly in urine have no obvious effect on the detection,indicating the well selectivity of the method.The spiked results obtained in real urine samples exhibit a recovery ranged from 91.4%to 109.2%.

关 键 词：8-羟基-2’-脱氧鸟苷 无标记检测 上转换荧光 纳米金 尿样 

分 类 号：X132[环境科学与工程—环境科学]