Background: Atopic dermatitis (AD) is a chronic inflammatory skin disease prevalent in children with a rate of 60–90% of superantigen producing staphylococcal colonisation that may play a role in its pathogenesis. Objective: This study aims to investigate the role of superantigen in the pathogenesis of atopic dermatitis by identifying enterotoxins producing S. aureus on the skin and nose of the patients using reverse passive latex agglutination test. Methodology: This study comprised 44 cases with atopic dermatitis attending the Outpatient Clinic of Dermatology & Paediatric Departments of Tanta University Hospital from December 2016 to the end of November 2017. Twenty healthy control volunteers were included. Results: Bacteriological skin cultures of patients revealed colonization with staph. aureus in(72.2)%, whereas, only 6 normal subjects (30%) showed positive cultures. This difference was statistically significant. Culture of nasal swabs revealed colonization with S. aureus in 21 atopic patients (47.2%) and only 8 subjects of control group (40%) with a statistically non significant difference. Wet lesions (in 23 patients) showed highly statistically significant colonization with S. aureus in comparison to dry lesions (in 6 patients). Out of the (32) positive staph. cultures from the skin of patients,(18) isolates were enterotoxin producers;(3) isolates produced enterotoxin A,(4) produced enterotoxin B,(9) produced enterotoxin C and (2) produced enterotoxin D. In control group, only (2) positive skin cultures were enterotoxins producers; (1) produced enterotoxin B and (1) produced enterotoxin C., the statistical difference was significant. Out of (21) nasal positive Staphylococcal cultures, 6 isolates were enterotoxins producers. Four of them produced enterotoxin B and two produced enterotoxin C. In control group, only 2 positive nasal cultures were enterotoxins producers; one produced enterotoxin B and one produced enterotoxin C. After treatment, skin lesions were clinically improved and the number of positive cultures were reduced from 18 to 8 (6 of them were still superantigen producing). This difference was significant. Conclusion: It is concluded that daily skin care, use of topical steroids and systemic anti-Staphylococcal antibiotics, are necessary to reduce skin inflammation.