We evaluated the reproducibility of atopy patch test reactions and the quality and quantity of itch in 16 patients with atopic eczema and a history of a positive atopy patch test reaction, comparing three different application sites. The allergen was re-applied simultaneously on both forearms and the back. Intensity and quality of pruritus were evaluated using a visual analogue scale and the Eppendorf itch questionnaire, respectively. The atopy patch test reaction was highly reproducible, occurring in 15/16 (94%) patients. Pruritus was reported by 14/16 (88%) patients. There was no significant difference in either the intensity or quality of itch between the two forearms and the back (p>0.05). The mean peak visual analogue scale itch score was comparable across all three test sites (range 28.3-31.9). Regarding quantification of test reactions, a positive reaction was more frequent on the back (94% versus 69% on the arms) and the peak atopy patch test score was higher on the back compared with the arms (right forearm, p=0.0018 and left forearm, p=0.0683). Allergens should preferably be applied on the back for the atopy patch test. However, the atopy patch test can induce atopic itch irrespective of the application site.