Development of a Test System Based on Recombinant GM6 Antigen from Trypanosoma evansi for the Determination of Surra in Horses
Akhmetsadykov N. Kydyrov T. Akhmetzhanova M. Akhmetova G. Berdikulov M.
2024ResearchersLinks Ltd
Journal of Animal Health and Production
2024#12Issue 11 - 10 pp.
Trypanosomosis, such as Surra, causes significant damage to livestock worldwide. The development of new serological diagnostic methods is highly relevant. The primary objective of this study was to produce a recombinant GM6 antigen derived from T. evansi and investigate its immunological properties for the detection and diagnosis of trypanosomosis. To create a recombinant antigen, bioinformatic analysis of the primary structure of T. evansi GM6 antigen from several regions was carried out. The protein sequence was reverse translated to the nucleotide sequence, after which the gene was synthesized using solid-phase method. The gene fragment, encoding the GM6 protein, was inserted or cloned into the pET28 expression vector after synthesis. The obtained sequences were checked by sequenc-ing for correspondence to the matrix molecule. For transformation E. coli BL21 (DE3) strain was used. To assess the efficacy of the antigen, immunoassay analysis and blot-hybridization techniques were employed. Obtained results showed that 1mM isopropyl ß-D-1-thiogalactopyranoside induction at 2℃ for 18 hours was the most optimal con-dition for expression of recombinant GM6 T. evansi antigen. From a 500 ml of growth medium, a total of 3.4 mg of recombinant protein was successfully obtained. A recombinant GM6 antigen of T. evansi encoding a 30 kDa polypep-tide was developed, which has diagnostically significant sensitivity in the enzyme immunoassay at a dilution of 1:6400 with sera of horses and donkeys infected with trypanosomosis. The proposed test system can be used for diagnosis of trypanosomosis in the early stages of the disease.
Amino Acid Sequence , Serological Detection , Trypanosomosis , Vector System , Veterinary Medicine
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Research and Production Enterprise “ANTIGEN”, Abay, Kazakhstan
Faculty of Water, Land and Forest Resources, Kazakh National Agrarian Research University, Almaty, Kazakhstan
Department of Biological Safety, Kazakh National Agrarian Research University, Almaty, Kazakhstan
Republican State Enterprise on the Right of Economic Management, National Reference Center for Veterinary Medicine, Committee for Veterinary Control and Supervision of the Ministry of Agriculture of the Republic of Kazakhstan, Astana, Kazakhstan
Research and Production Enterprise “ANTIGEN”
Faculty of Water
Department of Biological Safety
Republican State Enterprise on the Right of Economic Management
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