Homogeneous time-resolved fluorescence assay for the detection of ricin using an aptamer immobilized on europium-doped KGdF4 nanoparticles and graphene oxide as a quencher
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- 作者：Yukun Huang ; Xiujuan Chen ; Shijia Wu ; Nuo Duan ; Ye Yu ; Zhouping Wang
- 关键词：Time ; resolved fluorescence ; Graphene oxide ; Aptamer ; Ricin ; Fluorescence resonance energy transfer
- 刊名：Microchimica Acta
- 出版年：2015
- 出版时间：April 2015
- 年：2015
- 卷：182
- 期：5-6
- 页码：1035-1043
- 全文大小：1,375 KB
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- 刊物主题：Chemistry
Analytical Chemistry
Inorganic Chemistry
Physical Chemistry
Characterization and Evaluation Materials
Monitoring, Environmental Analysis and Environmental Ecotoxicology- 出版者：Springer Wien
- ISSN：1436-5073
文摘
We have developed an aptamer-based assay for the food toxin ricin. It is based on a competitive quenching strategy along with time-resolved fluorescence detection. The assay involves the following steps: (a) The aptamer is immobilized on the europium-doped KGdF4 nanoparticles (NPs); (b) these NPs are added to the sample where ricin binds to the aptamer; (c) graphene oxide (GO) is added and competitively binds to the aptamer on the NPs that are not blocked by ricin; this causes the quenching of the fluorescence of the NPs; (d) fluorescence is detected at 593 nm in a microplate reader in the time-resolved mode at an excitation wavelength of 273 nm, a delay time of 100 μs, and a gating time of 1 s. Under optimal conditions, the calibration plot is linearly related to the concentration of ricin in the 50 pg·mL? to 50 ng·mL? range (R2 = 0.9975), and the limit of detection is 8 pg·mL?. The method was compared to a standard ELISA, and correlation was excellent. The assay presented here provides a sensitive, dependable and convenient platform that is expected to have promising applications for the homogeneous assay of various other target analytes. Graphical Abstract A homogeneous time-resolved fluorescence assay was developed for the determination of ricin in portable water by FRET between KGdF4:Eu3+ nanoparticles-tagged aptamer and graphene oxide, with a good linearity and the limit of detection of 0.8?×-0?1?g?mL?.