Introduction:- There is always possibilities of transmission of transfusion transmissible diseases. Blood transfusion can transmit Human immunodeciency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV), through infected blood. Even Routine serological screening test and ELISA [Enzyme-linked Immunosorbent Assay] can not detect all infected blood units. Hence, to overcome this drawback and to minimise the risk, NAT [Nucleic Acid Testing] provides additional protection with higher sensitivity and specicity. Screening of Aim/Objective:- transfusion transmissible infections {HIV, Hepatitis B &C} using NAT technique to determine occurrence of seropostive blood donors in ELISA negative individuals. An observational study was conducted in Department of Pathol Methodology:- ogy Blood Bank, Gandhi Medical College and Associated Hospitals, Bhopal, where NAT investigation was conducted in all ELISA negative blood donors over the course of 1 year (04 Feb 21 to 04 Feb 22). 12- month data was compiled using Ms excel and analysed using Epi Statistical Analysis:- Result:- info 7.2. All the ELISA Negative(1271) blood units were tested by NAT. NAT was positive in 5 out of 1271 for HBV (0.4% cases). NAT was positive in 1 out of 1271 for HCV (0.1% cases). Dual testing of blood and blood products using high sensitivity serologic Conclusion:- al tests like NAT, aids in the detection of potentially infectious diseases in all stages of infection of HIV, HEPATITIS B and HEPATITIS C. In order to reduce overall treatment cost and boost effectiveness of healthcare system, it is advised to perform NAT in all ELISA negative blood donor samples.