Homogeneous fluorescent immunoassay for the simultaneous detection of chloramphenicol and amantadine via the duplex FRET between carbon dots and WS₂ nanosheets

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 WS₂ 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 WS₂ 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.