The aim of this study was to evaluate the role of serum human epididymis protein 4 (HE4) as a non-invasive biomarker for the diagnosis of liver fibrosis in children with chronic liver diseases (CLD). This case-control study was conducted at Benha University Hospital, Egypt, involving 60 children with CLD and 60 healthy children as a control group. HE4 levels were measured by ELISA and compared with liver biopsy results. The CLD group had significant higher HE4 (median: 110.7, IQR: 96.7–120.4 pmol/L) compared to control group (median 42.07, IQR: 41.67–43.05 pmol/L), p 48.3 pmol/L, HE4 diagnosed cases with mild fibrosis with a sensitivity of 95% and specificity of 91.3%. At a cutoff point >144.3 pmol/L, HE4 diagnosed cases with severe fibrosis with a sensitivity of 98% and specificity of 93.1%. Serum HE4 is a potential non-invasive marker for detecting liver fibrosis and its severity in children with CLD.