Background: While Staphylococcus aureus (S. aureus) colonization has been thoroughly studied in atopic dermatitis (AD), where S. aureus is related to flares and considered a trigger factor, S. aureus colonization in hand eczema (HE) has only been sparsely studied. Objectives: To examine the 1-week prevalence of S. aureus colonization in HE patients, and its association with severity, HE subtype, AD, and nasal S. aureus colonization compared with healthy controls. Methods: In a case-control study of 50 adult HE patients and 50 healthy controls, bacterial swabs from lesional skin (patients only), non-lesional skin (dorsal hand), and the nasal cavity were sampled for culturing of S. aureus on days 1, 3, 5 and 8. Participants were characterized by demographics, AD, HE subtype, filaggrin gene mutation status, and HE severity. Results: Twenty-seven HE patients (54%) were colonized with S. aureus on the hand compared to one control (2%) (P <.01). Nasal S. aureus colonization was found in 72% of patients and 22% of controls (P <.01). For patients, S. aureus colonization on the hands was associated with an atopic HE subtype and HE severity (P =.01 and P <.01, respectively). Conclusions: Both hand and nasal S. aureus colonization were highly prevalent among HE-patients and may have an impact on the persistence of HE.