In this study, we assessed the change in skin microbiota composition, relative abundance, and diversity with skin physiology disruption induced by SLS patch. Healthy women declaring to have a reactive skin were submitted to a 0.5% aqueous sodium lauryl sulfate solution application under occlusive patch condition for 24 h. Skin properties were characterized by tewametry, corneometry, and colorimetry and bacterial diversity was assessed by 16S rRNA sequencing. Analysis before and one day after SLS patch removal revealed an increase of skin redness and a decrease of stratum corneum hydration and skin barrier function. The relative abundance of taxa containing potential pathogens increase (Firmicutes: Staphylococcaceae; Proteobacteria: Enterobacteriaceae, Pantoea) while some of the most occurring Actinobacteria with valuable skin protection and repair capacities decreased (Micrococcus, Kocuria, and Corynebacterium). We observed an impaired skin barrier function and dehydration induced by SLS patch disturb the subtle balance of skin microbiota towards skin bacterial community dysbiosis. This study provides new insights on the skin bacterial composition and skin physiology simultaneously impaired by a SLS patch.