The present study aimed to assess the safety of a three-level stepwise oral food challenge (OFC) in patients with peanut allergy.This retrospective analysis reviewed laboratory test results and food allergy histories (obtained from medical records) of participants who underwent OFC using peanut butter for the first time at Miyagi Children's Hospital between January 2015 and December 2017. Total food dose was classified as follows: step 1, 0.1 g peanut butter; step 2, 0.5g peanut butter; and step 3, 3g peanut butter.A total of 114 participants were included. OFC-positive rates were 52% (step 1), 35% (step 2), and 20% (step 3). Of the 41 OFC-positive participants, 1 required an intramuscular adrenaline injection during step 3. Peanut allergy history rates and peanut- and Ara h 2-specific immunoglobulin E (sIgE) titers were significantly higher during the low-dose step.The three-level stepwise OFC with peanut butter is safe for patients with peanut allergy, as indicated by peanut- and Ara h 2-sIgE titers.