Current study measured the amplitude of low-frequency fluctuations (ALFF) and insular functional connectivity (FC) in Hepatitis C virus patients and compared with age and gender matched control using RESTplus software. HCV patients showed significantly reduced ALFF in the right insula. The right insular FC strength was lower in patients than control. Neuropsychological test scores were significantly lower in HCV patients. Changes in ALFF and FC provide new evidences of altered neuronal activity in HCV patients.