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Analytical methods with high sensitivities and short assay times are urgently required for the screening of "zero tolerance" hazardous substances in food. Herein, we propose a fluorescent immunoassay for the highly sensitive and rapid analysis of chloramphenicol (CAP) based on carbon dots (CDs)-encapsulated CaCO 3 nanospheres and magnetic nanoparticles (MNPs). The fluorescent immunoprobes were prepared by coupling the anti-CAP antibodies to carboxymethyl cellulose-functional CDs@CaCO 3 nanospheres. Chitosan-modified MNPs with "core-shell" structures were prepared and then conjugated to the CAP hapten, acting as the nano-carrier and interface for the immunoreaction. With the assistance of MNPs, the established fluorescent immunoassay achieved the sensitive detection of CAP in chicken with a limit of detection of 0.03 μg kg -1 and recoveries ranging from 83.7%-105.0%. The analysis results of the fluorescent immunoassay were evaluated by the enzyme-linked immunosorbent assay, having a correlation coefficient of 0.981. Our work provides a rapid, facile, and reliable strategy for the highly sensitive analysis of food contaminants based on "green" fluorescent nanoprobes., (Copyright © 2020 Elsevier B.V. All rights reserved.)