A novel recombinant Theileria annulata surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis
Ryskeldina A. Korobeinikov A. Tursunbay N. Berdikulov M. Shevtsov A. Bauer C. Mukhanbetkaliyev Y. Kuibagarov M.
August 2024Veterinary World
Veterinary World
2024#17Issue 81936 - 1942 pp.
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. Ryskeldina, et al.
cattle , indirect enzyme-linked immunosorbent assay , Kazakhstan , recombinant T. annulata surface protein , Theileria annulata
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National Center for Biotechnology, Astana, 01000, Kazakhstan
S. Seifullin Kazakh Agro Technical Research University, Astana, 010011, Kazakhstan
National Veterinary Reference Center, Astana, 010000, Kazakhstan
Institute of Parasitology, Justus Liebig University Giessen, Giessen, 35392, Germany
National Center for Biotechnology
S. Seifullin Kazakh Agro Technical Research University
National Veterinary Reference Center
Institute of Parasitology
10 лет помогаем публиковать статьи Международный издатель
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