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Herein, we proposed a duplex and homogeneous fluorescent immunoassay for the simultaneous detection of amantadine (AMD) and chloramphenicol (CAP) residue in chicken breast with both high sensitivity and short assay time. The immunoassay was based on the fluorescence resonance energy transfer (FRET) between hapten-labeled carbon dots (CDs) and antibody-modified WS 2 nanosheets. To achieve the duplex FRET, polyethyleneimine-functionalized blue and green emissive CDs with separated emission were synthesized via a one-pot hydrothermal method and directly coupled with the haptens of AMD and CAP, serving as the energy donors. The antibodies were modified on the surface of WS 2 nanosheets with high quenching efficiency to construct the energy acceptor. The specific immunoreaction could trigger the efficient FRET between the donors and the acceptors, causing the fluorescence quenching of CDs. The developed immunoassay was applied to simultaneously detect AMD and CAP, having the detection limit of 0.10 ng g -1 and 0.06 ng g -1 , respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Here we report a novel ultrasensitive surface-enhanced Raman scattering (SERS) immunosensor based on the flower-like gold nanoparticles (AuNFs) and magnetic bead separation for homogeneous detection of amantadine (AMD) in chicken just by one-step. The 5, 5'-dithiobis (2-nitrobenzoicacid) (DTNB) modified AuNFs and N-(1-adamantyl) ethylenediamine (AEDA) conjugated denatured BSA (AEDA-dBSA) was used as the SERS nanoprobe. And the capture probe was anti-AMD monoclonal antibody (mAbs)-functionalized magnetic beads (MNBs-mAbs). An immunoreaction occurred between free AMD and SERS nanoprobe for competing limited binding sites of MNBs-mAbs. This work combined inherent sensitive property of SERS with antibody-antigen highly specificity recognition for the AMD detection. The analytical results showed that the SERS-based immunosensor was sensitive, simple and reliable with a limit of detection (LOD) of 0.005 ng/mL for AMD, which were 2 orders of magnitude better than an enzyme-linked immunosorbent assay based on the same immunoreagents. Analysis of AMD-spiked chicken samples revealed that the developed immunosensor provided accepted recoveries ranging from 74.76%-89.34% with coefficient of variation less than 15.04%. This strategy represents a simple, reliable, and universal approach for detection of chemical contaminants in food samples., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Highlights • Electrostatic self-assembly of CDs@MnO 2 is used to develop a new fluorescent immunoassay. • The fluorescent immunoassay has significantly high sensitivity for the detection of amantadine in chicken.. • The alkaline phosphatase-dependent fluorescent immunoassay has a great potential in food analysis. Abstract Fluorescence immunoassays are rapid, convenient and cost-effective for the sensitive quantitation of chemical contaminants in foodstuff. In this study, a competitive fluorescence ELISA was developed for the sensitive detection of amantadine (AMD) based on the alkaline phosphatase (ALP)-triggered fluorescence "turn-on" signals. As a fluorescence substrate, carbon dots (CDs) were adsorbed onto the surface of the MnO 2 nanosheets (NSs) and formed a nanoassembly of p-CDs@MnO 2 NSs which results in the fluorescence quench of CDs. The ALP labelled on antibody could catalyze the hydrolysis of the 2-phosphoascorbic acid into ascorbic acid. The latter could then reduce and decompose the MnO 2 NSs, which was accompanied by the release of CDs from the surface of MnO 2 NSs and led to the fluorescence recovery of CDs. The change of the fluorescence intensity is related to the concentration of AMD in solution and thus could be applied to detect AMD in an ALP-based ELISA system. The fluorescent ELISA showed a linear detection for AMD in the range of 0.048 ng mL−1 to 1.1 ng mL−1 with a detection limit (LOD) of 0.035 ng mL−1. The novel fluorescent ELISA shows potential for the highly sensitive detection of AMD and other analytes in food analysis. [ABSTRACT FROM AUTHOR]