PurposeThis prospective study assessed the value of ultrasonography (US) in the evaluation of hepatosplenic fungal infections (HSFI).MethodsThirty‐two pediatric participants with confirmed onco‐hematological diseases and HSFI were included. Lesions in the liver and/or spleen were detected by US, magnetic resonance imaging (MRI), or computed tomography (CT).ResultsOf the participants, 11 (34%) had confirmed HSFI, while 21 (66%) had highly suspected HSFI. The US, CT, MRI, and fungal blood cultures demonstrated positive results in 31, 19, 25, and 7 patients, respectively. US had a significantly higher detection rate than CT, MRI, and fungal blood cultures (p < 0.05). The “bull's eye” phenomenon was a distinctive US feature of HSFI. Follow‐up examinations indicated that after a mean of 7.7 (1–15) months, liver and/or spleen lesions disappeared in five patients. The lesion was significantly smaller in 10 patients. Residual calcifications were detected in 15 patients. Two patients died.ConclusionConclusively, the US may substitute for tissue biopsy, other imaging modalities, or fungal blood culture for the confirmation of HSFI, and may guide better antifungal treatment, thus achieving better outcomes.