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Abstract Eczema and psoriasis are heterogeneous cutaneous inflammatory disorders with broad and occasionally overlapping diagnostic criteria, making it difficult to distinguish psoriasis from eczema. Despite the reality that potential biomarkers, such as CCL27 and NOS2 have been identified for differentiating psoriasis and eczema. The current study’s goals included determining the association between CCL27 and NOS2 expression level and disease severity as well as evaluating the expression level of CCL27 and NOS2 as a biomarker for distinguishing psoriasis and atopic dermatitis. This case-control study was carried out on 100 patients (50 with psoriasis and 50 with atopic dermatitis), and 50 age- and sex-matched controls were recruited after verbal consent was obtained. Patients who were receiving local or systemic treatment or who had an inflammatory skin disorder were excluded. Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) was used to evaluate the CCL27 and NOS2 expression level. Our study found a highly significant increase in CCL27 expression (P<0.00001) in eczema patients when compared with psoriasis and healthy control groups. To detect eczema, CCL27 expression levels with a cutoff value of 5.39-fold change had a high sensitivity (91.2%) and specificity (100%). In eczema patients, CCL27 was positively correlated with disease severity using SCORing Atopic Dermatitis scores |