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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. [ABSTRACT FROM AUTHOR]