With the increased utilization of assisted reproductive technology (ART), concerns about the potential health risks for ART-conceived babies have also been raised. Increased prevalences of allergic and metabolic diseases have been reported among ART offspring. This study aimed to evaluate the impact of IVF on the tendency to develop allergic responses following ovalbumin (OVA) sensitization in IVF-conceived mice. Mice were divided into four groups (non-OVA naturally conceived, OVA naturally conceived, non-OVA IVF-conceived, and OVA IVF-conceived). In the OVA groups, the mice were subjected to intraperitoneal and intranasal immunization with OVA. Two days after the final immunization, blood samples were taken, and the serum levels of IgE and IL-4 were detected by ELISA. The mice were sacrificed by cervical dislocation, their spleens and lungs were removed, and their weights were measured and recorded. Sensitization with OVA resulted in significantly increased concentrations of IL-4 and total IgE, as well as increased lung and spleen weights, among offspring from both natural and IVF conception. The concentrations of IgE and IL-4 and the lung and spleen weights in IVF-conceived mice were significantly higher compared to those in naturally conceived mice before and after sensitization with OVA. It is concluded that compared to naturally conceived mice, IVF-conceived mice exhibit a greater tendency to develop allergic responses against OVA.