Veterinary Research Forum
Vet Res Forum

Development of AMA-1 Antigen based ELISA kit for the diagnosis of Babesia infection in Cattle

Articles in Press

Document Type : Original Article

Authors

Bilal Aslam 1
Abdul Hafeez 2
Muhammad Shahid Mahmood 3
Faiza Aslam 4
Azhar Rafique 5
Sulaiman Fahad Aljasir 6

1 Department of Preventive veterinary Medicine, College of Veterinary Medicine, Qassim University Buraydah Saudi Arabia

2 Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan

3 Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan

4 Livestock and Dairy Development Department, Government of Punjab, Pakistan

5 Department of Zoology, Government College University, Faisalabad, Pakistan

6 Department of Veterinary Preventive Medicine, College of Veterinary Medicine, Qassim University Buraydah, Saudi Arabia

10.30466/vrf.2025.2065465.4829
Abstract
The current finding reports the development of iELISA with recombinant AMA-1 (rAMA-1) to identify an infection of Babesia in naturally infected cattle. The 48 kDa protein-encoding recombinant AMA-1 gene was cloned into the pET-28a (+) expression vector and expressed in E. 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; uninfected sera as well as Babesia positive samples with other species including B. bigemina, B. divergens, B. major, B. occultans 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 commercially available ELISA kit as the 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.

Keywords

Babesia
AMA-1 gene
ELISA
Domestic Cattle
Subjects
Veterinary Research Forum

Articles in Press, Accepted Manuscript
Available Online from 10 June 2026

  • Receive Date 10 July 2025
  • Revise Date 07 September 2025
  • Accept Date 11 October 2025

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us