Background Acute acalculous cholecystitis (AAC) is the most common form of acute cholecystitis in the pediatric population and some cases could have fatal complications. Epstein-Barr virus (EBV) is by far the most knowledgeable viral agent implicated in the pathogenesis of AAC in recent years. Thus, the risk factors in the development of AAC were favorably determined in the settings of EBV hepatitis. Methods Patients below 18 years of age, who had been admitted to Kaohsiung Chang Gung Memorial Hospital and were diagnosed with AAC concomitant with EBV hepatitis between January 2012 and March 2021, were retrospectively analyzed in our study. Our analyses comprised of risk factors including gestational age, age, gender, body weight, history of abdominal surgery, body temperature, duration of fever and hospitalization, clinical presentation, blood test parameters, sonographic findings, and medication. Results The results showed that the mean duration of fever, the presence of ascites, and increased atypical lymphocytes were statistically significant in patients with AAC, symbolizing a more severe disease burden as compared to those without AAC. The presence of ascites in sonographic findings and atypical lymphocytosis over 6.25% may be the two most significant predictive factors of developing AAC in children with EBV hepatitis. Conclusions Our study concluded that children with EBV-related AAC have presented with longer duration of fever, ascites, and atypical lymphocytosis over 6.25%. Further observation and close follow-ups may be suggested in high-risk children to prevent the occurrence of complications that require an emergent operation.