Trace detection of chiral molecules, which is of great significance in chemical, biological, medical and pharmaceutical sciences, requires microscopic techniques at the single-particle or single-molecule level. Although ensemble experiments show that the circular dichroism of chiral molecules can be amplified by plasmonic nanocrystals, trace detection of small chiral molecules remains challenging due to weak signals that are far below the detection limit. Herein, we demonstrate trace detection of chiral J-aggregated molecules adsorbed on individual Au nanorods (NRs) using single-particle circular differential scattering (CDS) spectroscopy. Through measuring the single-particle CDS spectra, we identified dip-peak bisignatures and further determined the chirality by matching them with calculations modelled with chiral media. We therefore find that plasmonic nanocrystals can dramatically amplify the circular dichroism of strongly coupled molecules to a detectable level so that the detection limit is as low as 3.9 × 103 molecules on an individual plasmonic nanoparticle, whereas 2.5 × 1012 molecules free in solution are barely detectable using a commercial circular dichroism instrument, suggesting a significant amplification factor of 108. Our method provides a promising strategy with a high amplification factor, shedding light on the trace detection of chiral molecules using optical microscopic methods.