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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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Abstract

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−1 to 50 ng·mL−1 range (R2 = 0.9975), and the limit of detection is 8 pg·mL−1. 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.

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 × 10−11 g mL−1.

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Acknowledgments

This work was partly supported by the National S&T Support Program of China (2012BAK08B01), S&T Supporting Project of Jiangsu Province (BE2011621), NCET-11-0663, and JUSRP51309A.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

    Yukun Huang, Xiujuan Chen, Shijia Wu, Nuo Duan & Zhouping Wang

  2. Zhangjiagang Entry-Exit Inspection and Quarantine Bureau, Zhangjiangang 215600, China

    Ye Yu

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  1. Yukun Huang
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  2. Xiujuan Chen
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  3. Shijia Wu
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  4. Nuo Duan
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  5. Ye Yu
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  6. Zhouping Wang
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Correspondence to Shijia Wu or Zhouping Wang.

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Huang, Y., Chen, X., Wu, S. et al. 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. Microchim Acta 182, 1035–1043 (2015). https://doi.org/10.1007/s00604-014-1422-3

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