Hepatitis B infection is a prominent infectious disease caused by hepatitis B virus (HBV), which infect liver and is considered as the main cause of liver cirrhosis, fibrosis and liver cancer worldwide. A pro-inflammatory cytokine Interleukin32 is believed to have a role in chronic HBV infections. Since its role in CHB infections is remain unclear, this study was done to detect IL-32 gene expression in CHB patients in order to identify its exact role. A total number of 110 blood samples were collected from Gastroenterology and Hepatology Teaching Hospital in Baghdad Medical City from CHB patients for both males and females with different age groups according to the research ethics form then sent to Central Public Health Laboratory (CPHL), National AIDS and Viral Hepatitis center, Baghdad, Iraq between October 2020 and March 2021. The immunological part for evaluation of HBsAg (HB surface antigen) and HBcAb IgG (HB core antibody IgG) was accomplished by Enzyme Linked Immunosorbent Assay technique (ELISA) while, the molecular part of this study was achieved by reverse transcription- quantitative PCR (RT-qPCR) to detect the gene expression of Interleukin-32 in Chronic HBV patients. The results showed a decrease in IL32 expression in most of the studied samples of Iraqi chronic HBV patients (69.73%), while only (30.26%) of them are showed increase in IL-32 expression. Statistical analysis ensured that differences among age groups of both gender are not significant.