Background: Although there are several case reports of wet wipe-associated contact dermatitis, the prevalence of wipes as a source of allergic contact dermatitis in larger populations and the responsible allergens are largely unknown. Objective: The aim of the study was to determine the prevalence of wet wipes as a source of contact allergy and the most commonly associated allergens in a North American tertiary referral patch test population. Methods: Data collected from 2011 to 2014 by the North American Contact Dermatitis Group was used to conduct a retrospective cross-sectional analysis of patient demographics and patch test results associated with the triple-digit source code for "wet wipe." Results: Of the 9037 patients patch tested during the study period, 79 (0.9%) had a positive patch test reaction to an allergen identified with a wet wipe source. The most commonly associated allergens were preservatives, including the following: methylisothiazolinone (MI) (59.0%), methylchloroisothiazolinone (MCI)/MI (35.6%), bronopol (2-bromo-2-nitropropane-1,3-diol) (27.4%), and iodopropynyl butylcarbamate (12.3%). Fragrance (combined) represented 12.3%. Anal/genital dermatitis was 15 times more likely (P < 0.0001) in those with wet wipe allergy. More than 92% of patients with wipe-associated contact allergy had their contact allergens detected by the North American Contact Dermatitis Group screening series. Conclusions: Wet wipes are an important source of contact allergy. Preservatives are the main allergens, especially isothiazolinones.