Optimization of laboratory animals hyperimmunization schemes with a highly purified rabies virus antigen

Mukhamedzhanova, A. G.; Ахмадеев, Р. М.; Miftakhov, N.R.; Nasyrov, Sh.M.; Aleeva, Z. Z.; Yarullina, G.M.; Arutyunjan, G.S.

doi:10.30896/0042-4846.2020.23.10.25-29

"Veterinary Medicine" Journal

2020

DOI: 10.30896/0042-4846.2020.23.10.25-29

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Optimization of laboratory animals hyperimmunization schemes with a highly purified rabies virus antigen

A. G. Mukhamedzhanova¹,

Р. М. Ахмадеев²,

N.R. Miftakhov³

et al.

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Construction of prokaryotic system for expression of porcine circovirus type 2 ORF-2 gene fragment

Galeeva,

Akhunova,

Usoltsev

et al. 2024

Veterinariâ segodnâ

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Porcine circovirus-associated diseases (PCVDs) are among the most significant challenges for pig farming in developed countries. Porcine circovirus type 2 (PCV-2) is considered the main etiological agent of postweaning multisystemic wasting syndrome in piglets. Mass PCVD occurrence has been reported in most regions of the world, that results in serious economic consequences. Optimal PCVD prevention is known to be achieved through a set of veterinary and sanitary measures in combination with vaccination. High evolutionary virus variability facilitating new genotype and strain emergence requires development of new candidate recombinant vaccines against PCV-2 infection. The study was aimed at construction of prokaryotic system for PCV-2 ORF-2 gene fragment expression and its functionality assessment. A genetic insert constructed from the most immunogenic type-specific PCV-2 epitopes based on genotype 2a, 2b, 2d strain and isolate consensus sequence was cloned into the expression vector pET-22b(+) that was incorporated into the Escherichia coli strain Rosetta 2(DE3). The transformants were selected based on the marker gene of ampicillin resistance on a selective medium. Target gene expression was induced by adding of isopropyl-β-D-1-thiogalactopyranoside at different concentrations. As a result, Escherichia coli Rosetta 2(DE3)/pET-22b-ORF-2 strain, a producer of capsid protein fragment (92–233 amino acid residues), was constructed. It was found that in the presence of 1 mM isopropyl-β-D-1-thiogalactopyranoside, the expression level of soluble truncated rCap was 35–40 mg/L 6 hours after induction. The expression product was tested for its specificity with indirect ELISA using whole-virion PCV-2-hyperimmunized porcine serum. It was shown that the positivity coefficient of producer strain cell lysates averaged to 4.34 (p < 0.005). The recombinant rCap protein is suitable for serological diagnosis and is also of interest as a vaccine component, which is the goal of our further studies.

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Construction of prokaryotic system for expression of porcine circovirus type 2 ORF-2 gene fragment

Galeeva,

Akhunova,

Usoltsev

et al. 2024

Veterinariâ segodnâ

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An immunochromatographic test for the detection of anti-tuberculosis antibodies

Khammadov,

Khaertynov,

Akhmadeev

et al. 2023

jour

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Tuberculosis is a dangerous socially significant disease of various animal species. According to Rosselkhoznadzor data for 2022, the situation with bovine tuberculosis is endemic, stable, long-term trends are decreasing, epidemic thresholds for ill health and incidence have not been overcome. It should be noted that in 2022, bovine tuberculosis was not officially detected, but this infection was registered in several pigs and wild boars. Take a point that the reduction in the number of cases of tuberculosis in cattle is decreased (to zero), the overall strategy for combating this disease is successful, but this does not mean that the need to develop new tests for the accelerated diagnosis of this infection has completely disappeared. The safety of service personnel directly depends on minimizing the risk of infection with pathogens common to humans and animals, one of which is tuberculosis. The fastest way to indicate infectious agents is by immunochromatographic analysis. This publication is devoted to the details that it is desirable to focus on when developing a diagnostic test based on immunochromatographic analysis, using the example of bovine tuberculosis. As an antigen in the presented study, native antigens of Mycobacterium bovis grown on a nutrient medium Levenshtein-Jensen with proven antigenic activity were used. Conjugation of antispecies antibodies was carried out with a colloidal solution of gold, (diameter of particle is 25±0,9 nm). In the course of the work, the optimal conditions for the manufacture of a diagnostic test for the detection of anti-tuberculosis antibodies by immunochromatographic analysis were shown.

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Optimization of laboratory animals hyperimmunization schemes with a highly purified rabies virus antigen

Cited by 2 publications

References 0 publications

Construction of prokaryotic system for expression of porcine circovirus type 2 ORF-2 gene fragment

Construction of prokaryotic system for expression of porcine circovirus type 2 ORF-2 gene fragment

An immunochromatographic test for the detection of anti-tuberculosis antibodies

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