Background: Amphotericin B is the broadest-spectrum polyene group antifungal agent. In the presence of an infection caused by a fungal agent resistant to alternative antifungal drugs, desensitization may be the only option to continue treatment. However, there is insufficient information and consensus regarding Amphotericin B desensitization protocols in the pediatric age group. Objective: In this study, we present our experience with five cases where successful desensitization protocols were applied with Liposomal Amphotericin B (AmBisome®; LAMB), along with a review of the literature on pediatric cases. We also provide a sample desensitization protocol that we successfully applied. Methods: In this retrospective study, pediatric patients who continued their treatment with the successful rapid desensitization protocol conducted at the Pediatric Allergy and Immunology Clinic of the Ministry of Health Ankara City Hospital between September 2019 and September 2023 were examined. In our study, desensitization protocols were applied based on Castell et al.’s desensitization protocol. Results: Between 2019 and 2023, five patients aged between 5 and 12 years, including three with hematologic malignancies, one with tuberculosis meningitis, and one with chronic granulomatous disease, were referred to us due to the development of anaphylaxis during their treatment with LAMB. Anaphylaxis is diagnosed clinically, considering the European Academy of Allergy and Clinical Immunology (EAACI). A 16-step desensitization protocol was prepared using LAMB solutions at four different dilutions (0.001 mg/ml, 0.01 mg/ml, 0.1 mg/ml, 1 mg/ml). Each solution consisted of four steps with a 15-minute infusion for each step. Patients was premedicated with 1 mg/kg/dose methylprednisolone and an antihistamine. Conclusion: The data we present on the successful application of a sample protocol to five cases, particularly in a pediatric setting, is a significant and valuable contribution to the field. This can provide important insights and potentially serve as a reference for medical professionals working with similar cases in the future.