Tehran University of Medical Sciences Journal of Medical Bacteriology 2251-8649 12 2 2024 04 22 34 42 Faeze Beik Mahdavi MSc, Islamic Azad University, Chalus Branch Department of medical science, Chalous, Iran Haniyeh Bashizadehfakhar Department of Human Geneticse, Science And Research Branch, Branch, Islamic Azad University, Tehran, Iran Melika Jalalian Department of cell and molecular sciences, faculty of advanced sciences & technology, Tehran medical science, Islamic Azad University, Tehran, Iran Shaghayegh Rangraz MSc, Islamic Azad University, Chalus Branch Department of medical science, Chalous, Iran 2024 01 10 2024 04 04 Background:   Since HCV virus is the primary cause of liver cirrhosis and cancer, prompt diagnosis and timely treatment of this disease can prevent many complications. Due to the importance and necessity of this study, it aims to comparatively evaluate RT-PCR and ELISA methods in order to detect the presence of HCV infection in patients admitted to Baghiyatallah Hospital. Methods:   Sera of 49 patients admitted to Baghiyatallah Hospital since September, 2019 to August, 2020, were tested for RNA detection of HCV virus using RT-PCR and for the presence of anti-HCV antibody at the same time. Results:   In this study, the mean age of patients was approximately 38.3+6.3. The percentage of positive cases of HCV virus in the studied patients according to ELISA test and PCR test were 28.6% and 20.4%, respectively. Percentage of HCV positive cases had p = 0.001 based on PCR and ELISA tests by age group, number of sexual partners, history previous HCV infection, liver cirrhosis, addiction and treatment of previous HCV infection which was significantly different; However, it was not significant in terms of gender (p = 0.5232). According to Kappa coefficient, the percentage of agreement is 91.8 in both methods which indicates that the two models are consistent (P = 0.001) and the diagnostic value of ELISA versus PCR with sensitivity and specificity were 100% and 89.7%, respectively. Conclusion:   ELISA susceptibility to anti-HCV antibody is more than 99%, but its specificity is low compared to RT-PCR method. On the other hand, these tests show only hepatitis C affection and does not distinguish between chronic acute or improved infection. It is not able to detect patients in the window phase, so the simultaneous application of ELISA and molecular methods is recommended to diagnose and follow the treatment of HCV virus. https://jmb.tums.ac.ir/index.php/jmb/article/view/511