Objectives: This study aimed to investigate the spectrum of bacterial pathogens and their antimicrobial sensitivity in pediatric patients admitted with febrile neutropenia (FN) and hematologic malignancies. Methods: This prospective, cross-sectional study was carried out on 65 children (60% boys, mean age 7.3 5.3 years) with hematologic malignancies and FN hospitalized at the Amir Hematology and Oncology Hospital, Shiraz, Iran, between December 2018 and May 2019. Samples were taken from blood, nose, axilla, and inguinal area (NAI). NAI cultures were repeated after 72 hours of admission, and blood culture was repeated if needed. A demographic questionnaire on age, sex, and admission time was filled out for each patient. Results: The most common grown bacteria in the NAI samples were coagulase-negative staphylococci (CoNS). Second cultures from these sites revealed nosocomial contamination. Eighteen patients (27.7%) had positive blood cultures, with 61.1 % of the isolated pathogens being Gram-negative (mostly Escherichia coli and Pseudomonas aeroginosa) and 38.9% Gram-positive. The blood cultures results did not match those of the NAI cultures in 61.1% of the patients. About one-fourth of the Gram-negative pathogens isolated from blood were resistant to cefepime, the main empirical antibiotic for FN management at our center. Conclusions: Gram-negative bacteria are predominant organisms in pediatric FN patients at our center, most resistant to cefepime. If a similar antimicrobial sensitivity pattern is observed in future studies, a change in local guidelines might be considered for FN management.