In the developing countries including Pakistan, hepatitis B virus (HBV) and hepatitis C virus (HCV) cause serious health problems. A range of risk factors are responsible for this infectious disease's spread. This study was carried out to detect virus type in suspected patients from Islamabad and Karachi (Southern and Northern parts of Pakistan) and evaluate the viral association with gender. Identification of viral hepatitis type was done by real time PCR (Cepheid, USA). Association of Hepatitis viruses (Hepatitis B & C) infection with gender was analyzed, among suspected patients reported in a Local Hospital of Karachi and Islamabad. A total of 916 suspected patients were screened for Hepatitis B (n=282) and Hepatitis C (n=634). Total pervasiveness of Hepatitis B and Hepatitis C was found to be more common in male by percentage analysis but statistical analysis results differ from these. Further studies are needed to characterize HBV and HCV viruses, causes of their spread and transmission with reference to Pakistan.