Vet World Vol.17 August-2024 Article - 32
Research Article
Veterinary World, 17(8): 1936-1942
https://doi.org/10.14202/vetworld.2024.1936-1942
A novel recombinant Theileria annulata surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis
2. S. Seifullin Kazakh Agro Technical Research University, 010011, Astana, Kazakhstan.
3. National Veterinary Reference Center, 010000, Astana, Kazakhstan.
4. Institute of Parasitology, Justus Liebig University Giessen, 35392, Giessen, Germany.
Background and Aim: Theileria annulata infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant T. annulata surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis.
Materials and Methods: Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in Escherichia coli strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of T. annulata-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for T. annulata and/or Theileria orientalis from southern Kazakhstan.
Results: The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of T. annulata. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of T. annulata PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with T. orientalis.
Conclusion: Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and T. annulata seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries.
Keywords: cattle, indirect enzyme-linked immunosorbent assay, Kazakhstan, recombinant T. annulata surface protein, Theileria annulata.
How to cite this article: Ryskeldina A, Korobeinikov A, Tursunbay N, Berdikulov M, Shevtsov A, Bauer C, Mukhanbetkaliyev Y, and Kuibagarov M (2024) A novel recombinant Theileria annulata surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis, Veterinary World, 17(8): 1936-1942.
Received: 2024-04-24 Accepted: 2024-07-25 Published online: 2024-08-27
Corresponding author: E-mail:
DOI: 10.14202/vetworld.2024.1936-1942
Copyright: Ryskeldina, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
