Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 16 3 2024 06 19 357 365 Hassan Seyyedhamzeh Department of Microbiology, Faculty of Basic Sciences, Jahrom Branch, Islamic Azad University, Jahrom, Iran Safar Farajnia Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Mohammad Kargar Department of Microbiology, Zand Institute of Higher Education, Shiraz, Iran Behzad Baradaran Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Farshid Kafilzadeh Department of Microbiology, Faculty of Basic Sciences, Jahrom Branch, Islamic Azad University, Jahrom, Iran 2023 08 19 2024 03 02 Background and Objectives: Helicobacter pylori is known as the main cause of gastrointestinal diseases including gastritis, gastric ulcer and stomach cancer. Serodiagnosis of H. pylori infection is a noninvasive and rapid method but the efficiency of this method is highly dependent to the antigens used. This study evaluated the efficacy of recombinant UreB-Omp18 and FliD for serodiagnosis of H. pylori infection. Materials and Methods: The genes encoding for fliD, ureB, and omp18 was amplified by PCR and cloned into pET-22b and pET-28a vectors. The constructs were expressed in E. coli BL21 and purified by affinity chromatography. The antigenic properties and diagnostic potential of the recombinant proteins were analysed by immunoblotting and ELISA, respectively. Results: The recombinant UreB-Omp18 and FliD with molecular weights of 48 kDa and 25 kDa were observed on SDS-PAGE and purified by the Ni-NTA column. The ELISA results showed that the sensitivity and specificity of recombinant UreB-Omp18 protein in serodiagnosis of H. pylori infection were 89% and 83%, respectively. Also, the sensitivity and specificity of the recombinant FliD protein were calculated to be 91% and 76%, respectively. Conclusion: The results indicated that the recombinant UreB-Omp18 and FliD could diagnose H. pylori infection with high sensitivity and specificity. https://ijm.tums.ac.ir/index.php/ijm/article/view/4364 https://ijm.tums.ac.ir/index.php/ijm/article/download/4364/1677