• 文献标题:   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
  • 文献类型:   Article
  • 作  者:   HUANG YK, CHEN XJ, WU SJ, DUAN N, YU Y, WANG ZP
  • 作者关键词:   timeresolved fluorescence, graphene oxide, aptamer, ricin, fluorescence resonance energy transfer
  • 出版物名称:   MICROCHIMICA ACTA
  • ISSN:   0026-3672 EI 1436-5073
  • 通讯作者地址:   Jiangnan Univ
  • 被引频次:   6
  • DOI:   10.1007/s00604-014-1422-3
  • 出版年:   2015

▎ 摘  要

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 mu s, 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 center dot mL(-1) to 50 ng center dot mL(-1) range (R-2 = 0.9975), and the limit of detection is 8 pg center dot 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.