Background: there is an increasing interest in using microRNAs (miRNAs) as biomarkers in liver disease. Diagnostic biomarkers in hepatitis C serve as a great benefit for the early treatment of HCV. Aim of the Work: the aim of this study was to evaluate the expression of miRNA-16 in Egyptian patients infected with HCV and their relation to different biochemical and clinicopathological parameters. Patients and Methods: twenty-five subjects were chosen, 16 HCV infected and 9 healthy controls. Collection, processing and storage of serum for evaluation of miRNA16 using RT-qPCR was done. We evaluated the power of miRNA as a diagnostic tool using ROC curve analysis. The prognostic significance of the investigated parameters in HCV patients was explored. Results: there is a highly significant difference of miRNA 16 expression between HCV patients and healthy controls. miRNA 16 has great sensitivity and specificity for differentiating between patients with HCV and healthy controls. Conclusion: miRNA 16 can be used as a potential diagnostic biomarker for HCV. In addition, it could be used for staging of the disease.