Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
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A combined application of molecular docking technology and indirect ELISA for the serodiagnosis of bovine tuberculosis

  • Song, Shengnan (College of Animal Science and Technology, Shihezi University) ;
  • Zhang, Qian (State Key Laboratory for Sheep Genetic Improvement and Healthy Production, Xinjiang Academy of Agricultural and Reclamation Science) ;
  • Yang, Hang (College of Animal Science and Technology, Shihezi University) ;
  • Guo, Jia (College of Animal Science and Technology, Shihezi University) ;
  • Xu, Mingguo (College of Animal Science and Technology, Shihezi University) ;
  • Yang, Ningning (College of Animal Science and Technology, Shihezi University) ;
  • Yi, Jihai (College of Animal Science and Technology, Shihezi University) ;
  • Wang, Zhen (College of Animal Science and Technology, Shihezi University) ;
  • Chen, Chuangfu (College of Animal Science and Technology, Shihezi University)
  • Received : 2021.10.19
  • Accepted : 2022.04.18
  • Published : 2022.05.31
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Abstract

Background: There is an urgent need to find reliable and rapid bovine tuberculosis (bTB) diagnostics in response to the rising prevalence of bTB worldwide. Toll-like receptor 2 (TLR2) recognizes components of bTB and initiates antigen-presenting cells to mediate humoral immunity. Evaluating the affinity of antigens with TLR2 can form the basis of a new method for the diagnosis of bTB based on humoral immunity. Objectives: To develop a reliable and rapid strategy to improve diagnostic tools for bTB. Methods: In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in Escherichia coli. The two antigenic proteins, MPT70 and MPT83, which were most valuable for serological diagnosis of bTB were screened. Molecular docking technology was used to analyze the affinity of MPT70, MPT83, dominant epitope peptide of MPT70 (M1), and dominant epitope peptide MPT83 (M2) with TLR2, combined with the detection results of enzyme-linked immunosorbent assay to evaluate the molecular docking effect. Results: The results showed that interaction surface Cα-atom root mean square deviation of proteins (M1, M2, MPT70, MPT83)-TLR2 protein are less than 2.5 A, showing a high affinity. It is verified by clinical serum samples that MPT70, MPT83, MPT70-MPT83 showed good diagnostic potential for the detection of anti-bTB IgG and M1, M2 can replace the whole protein as the detection antigen. Conclusions: Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.

Keywords

Acknowledgement

This research was supported by Open Fund of National Natural Key Research and Development Plan (No. 2017YFD0500304), Key R&D Project of Hebei Province (21322912D) and Corps Major Scientific and Technological Projects (No. 2017AA003).

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