College of Animal Science and Technology, Shihezi University, Shihezi 832000, China.
Collaborative Innovation Center for Prevention and Control of High Incidence Zoonotic Infectious Diseases in Western China, Shihezi 832003, China.
J Vet Sci. 2022 May;23(3):e50. doi: 10.4142/jvs.21270.
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.
To develop a reliable and rapid strategy to improve diagnostic tools for bTB.
In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in . 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.
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.
Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.
为了应对全球范围内不断上升的牛结核病（bTB）流行，迫切需要寻找可靠和快速的牛结核病诊断方法。Toll 样受体 2（TLR2）识别 bTB 的成分，并启动抗原呈递细胞介导体液免疫。评估抗原与 TLR2 的亲和力可以为基于体液免疫的 bTB 诊断新方法奠定基础。
开发一种可靠和快速的策略，以改进 bTB 的诊断工具。
本研究在. 中表达和纯化了 16 种 bTB 特异性重组蛋白。筛选出对 bTB 血清学诊断最有价值的两种抗原蛋白 MPT70 和 MPT83。采用分子对接技术分析 MPT70、MPT83、MPT70 优势表位肽（M1）和 MPT83 优势表位肽（M2）与 TLR2 的亲和力，并结合酶联免疫吸附试验检测结果评价分子对接效果。
结果表明，蛋白（M1、M2、MPT70、MPT83）-TLR2 蛋白的相互作用表面 Cα-原子均方根偏差小于 2.5A，表现出高亲和力。临床血清样本验证表明，MPT70、MPT83、MPT70-MPT83 对检测抗-bTB IgG 具有良好的诊断潜力，M1、M2 可替代整个蛋白作为检测抗原。
分子对接评估 bTB 蛋白与 TLR2 的亲和力并结合 ELISA 为 bTB 的诊断提供了新的思路。
