BACKGROUND: Hepatitis B virus (HBV) is the most widespread and the most important type among hepatitis viruses, affecting about 350 million people globally. The present study was done to determine the common clinical features, serological and molecular parameters in the diagnosis of Hepatitis B Virus infection in clinically suspected patients. MATERIALS & METHODS: This study was carried out in the department of Microbiology and Molecular Diagnostics in association with the Department of Pathology at Krsnaa Diagnostics Ltd, Guwahati, Assam over a period of 3 months from May, 2024 till July, 2024. This study was conducted among 296 patients, who were clinically suspected for HBV infection and screened for Hepatitis B ICT (Hepa Card) serological test as well as HBV real time PCR molecular test with viral count quantification. The collected data was entered into MS excel followed by the analysis using SPSS version 21 (licensed to KDL). RESULTS: In our study, we noted that, out of a total of 296 HBV suspected patients, 163(55%) patients were in the age group of 20-30 years, followed by 71 (24%) patients were in the age group of 30-40 years, followed by 50(17%) patients were in the age group of 40-50 years, followed by 12(4%) patients were in the age group of 13-19 years. Out of total 296 patients, 185 came out to be reactive by rapid ICT card test and 111 came out to be HBV negative. 133 Came out to be reactive by real time PCR and 163 came out to be negative. Out of total 296 samples, samples which came out to be HBV reactive by ICT Card test but not by PCR was 87. And samples which came out to be HBV reactive by PCR but not reactive by ICT rapid card test were 39. The PCR results average log value was 5.07548 & average CT value was 26.80.