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Objective To explore the effect of empirical treatment and diagnostic-driven treatment in invasive fungal infection (IFD) during chemotherapy in children with acute lymphoblastic leukemia (ALL). Methods The data of 106 children with ALL who received antifungal treatment in pediatric hematology department of Sun Yat-Sen Memorial Hospital of Sun Yat-sen University from February 2018 to December 2020 were retrospectively analyzed. According to the treatment methods, they were divided into experience group (45 cases) and diagnosis group (61 cases). The incidence of IFD, mortality, fever time, medication time, adverse reactions and transfer to pediatric intensive care unit (PICU) were compared between the two groups. Results The incidence of IFD in the diagnosis group was higher than that in the experience group, there was significant difference (P <0.05). There were no significant differences in IFD related mortality and transfer to PICU between the two groups (P >0.05). There were significant differences in fever time before drug application and drug application time between the two groups (P <0.05). There were no significant differences in drug replacement and adverse reactions between the two groups (P >0.05). Conclusion Experience therapy and diagnostic-driven therapy are effective methods for the treatment of IFD in children with ALL during chemotherapy, which should be selected according to the specific situation of children in clinic. [ABSTRACT FROM AUTHOR]