Significance Optical chirality by light–matter interaction is a powerful strategy for discriminating the chirality of materials in modern physics. Here we present a vortex-dependent approach to yield a direct chiroptical response with monochromatic light. The chiroptical signal of ∼120% is obtained on intrinsically chiral microstructures by optical vortices in the visible range. The chiroptical properties of microstructures can be flexibly controlled by tuning their geometric features. We further demonstrate the strong chiral light–matter interaction by optical vortices for sensitively detecting optical chirality of molecules.