The nature of dark matter remains unknown to date, although several candidate particles are being considered in a dynamically changing research landscape1. Scalar field dark matter is a prominent option that is being explored with precision instruments, such as atomic clocks and optical cavities2–8. Here we describe a direct search for scalar field dark matter using a gravitational-wave detector, which operates beyond the quantum shot-noise limit. We set new upper limits on the coupling constants of scalar field dark matter as a function of its mass, by excluding the presence of signals that would be produced through the direct coupling of this dark matter to the beam splitter of the GEO600 interferometer. These constraints improve on bounds from previous direct searches by more than six orders of magnitude and are, in some cases, more stringent than limits obtained in tests of the equivalence principle by up to four orders of magnitude. Our work demonstrates that scalar field dark matter can be investigated or constrained with direct searches using gravitational-wave detectors and highlights the potential of quantum-enhanced interferometry for dark matter detection. © 2021, The Author(s).