Background: Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide, ranging from simple steatosis and non-alcoholic steatohepatitis (NASH) to cirrhosis, with its complications including hepatocellular carcinoma (HCC). The non-invasive diagnostic markers of NAFLD are gaining interest due to the invasive nature and potential side effects of liver biopsy, and are the current gold standard of diagnosis. Objective: This study aimed to evaluate hyaluronic acid (HA) as a potential non-invasive biomarker for diagnosis and prognosis of NAFLD and to compare it with the traditional non-invasive techniques. Patients and methods: This study included 63 subjects divided into 3 groups; including 21 patients with fatty liver, 21 patients with NASH, in addition to 21 healthy controls. Non-invasive assessment of liver fibrosis was done to all study subjects using aspartate aminotransferase to platelet ratio index (APRI) and fibrosis-4 score (FIB4), in addition to measurement of HA serum levels. Results: As regard assessment of liver fibrosis, there was a highly significant statistical difference between groups as regard APRI and Fib4 scores. As for HA there was a highly significant statistical difference between study groups. HA receiver operating characteristics curves (ROC) achieved excellent diagnostic performance for fatty liver with 100% sensitivity and specificity at cutoff of 25 ng/ml for fatty liver, versus a sensitivity of 95.2% and specificity of 85.7% for NASH at cutoff of 110 ng/ml. Conclusion: HA can be used as an accurate and specific non-invasive biomarker for the diagnosis of NAFLD and staging of the severity of disease compared with the traditional known non-invasive scores.