Faculty of Veterinary Medicine, Urmia University Veterinary Research Forum 2008-8140 17 6 2026 06 01 Development of apical membrane antigen-1 based ELISA kit for the diagnosis of Babesia infection in cattle 399 405 736498 10.30466/vrf.2025.2065465.4829 EN Muhammad Shahid Mahmood Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan Azhar Rafique Department of Zoology, Faculty of Life Sciences, Government College University, Faisalabad, Pakistan 0000-0001-8147-8820 Faiza Aslam Livestock and Dairy Development Department, Government of Punjab, Punjab, Pakistan Mian Abdul Hafeez Department of Parasitology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan Bilal Aslam Department of Veterinary Preventive Medicine, College of Veterinary Medicine, Qassim University, Buraydah, Saudi Arabia 0000-0002-5568-5049 Sulaiman Fahad Aljasir Department of Veterinary Preventive Medicine, College of Veterinary Medicine, Qassim University, Buraydah, Saudi Arabia Journal Article 2025 07 10 The current finding reports the development of immunocapture-enzyme linked immunosorbent assay (iELISA) with recombinant apical membrane antigen-1 (rAMA-1) to identify an infection of <em>Babesia </em>in naturally infected cattle. The 48.00 kDa protein-encoding rAMA-1 gene was cloned into the pET-28a (+) expression vector and expressed in Escherichia coli. The resulting congregate protein was refined under native settings. Towards the evaluation of the diagnostic potential of AMA-1 as a sero-diagnostic reagent, a panel of sera samples from Babesia infected cattle and uninfected sera, as well as <em>Babesia </em>positive samples with other species, including <em>B. bigemina</em>, <em>B. divergens</em>, <em>B. major</em>, <em>B. occultans</em>, were utilized. Additionally, the efficacy of rAMA-1-based serological assays was compared with commercially available kits using 200 samples taken from cattle suspected of babesiosis. The results demonstrated that the iELISA using rAMA-1 exhibited a diagnostic sensitivity of 88.89%, when compared to the commercially available ELISA kit as a reference test. The specificity of this assay was 76.66%. These findings suggest that the iELISA employing rAMA-1 can be utilized on large-scale epidemiological surveys and clinical detection of Babesia infection in cattle. https://vrf.iranjournals.ir/article_736498_c0a28496b3feb404623240960207ebd2.pdf