Key Lab of Environment and Health, School of Public Health, Xuzhou Medical University, Xuzhou, 221004, China.
School of Public Health, Jilin University, Changchun, 130021, China.
Biochem Biophys Res Commun. 2021 Feb 12;540:37-41. doi: 10.1016/j.bbrc.2020.12.098. Epub 2021 Jan 9.
Currently, brucellosis is a reemerged zoonotic infectious disease with an increased incidence in recent years. A simple, rapid and sensitive method for diagnosing brucellosis can help to reduce medical burden and economic loss. Previously, a multiple epitope recombinant protein was constructed based on linear B-cell epitope prediction tools. In this study, the recombinant protein was used as an antigen to study the immune response produced by immunized mice, and goat serum was used to verify its diagnostic accuracy. The production of antibodies was successfully induced in the vaccinated mice. Flow cytometric analysis revealed that the percentage of CD4, CD8 and the CD4+/CD8+ ratios were increased by T cell subsets in mouse splenocytes, indicating that the recombinant protein induced a strong immune response had strong immunoreactivity. Using indirect ELISA, the recombinant protein correctly diagnosed positive and negative brucellosis samples. Compared with the whole bacterial antigen, the recombinant protein had a weaker sensitivity but a stronger specificity. Animal experiments showed that the recombinant protein had good antigenicity, and indirect ELISA indicates that it can be used as an antigen to diagnose brucellosis. Therefore, the recombinant protein is a potential candidate antigen for brucellosis vaccine development and serological diagnosis.
目前，布氏杆菌病是一种重新出现的人畜共患传染病，近年来发病率有所上升。一种简单、快速、敏感的布氏杆菌病诊断方法有助于减轻医疗负担和经济损失。此前，基于线性 B 细胞表位预测工具构建了一种多表位重组蛋白。本研究以该重组蛋白为抗原，研究免疫小鼠产生的免疫应答，并用羊血清验证其诊断准确性。免疫小鼠成功诱导产生了抗体。流式细胞术分析显示，小鼠脾细胞中 T 细胞亚群的 CD4、CD8 和 CD4+/CD8+比值增加，表明重组蛋白诱导了强烈的免疫应答，具有很强的免疫原性。间接 ELISA 结果表明，该重组蛋白可正确诊断阳性和阴性布氏杆菌病样本。与全菌抗原相比，重组蛋白的敏感性较弱，但特异性较强。动物实验表明，该重组蛋白具有良好的抗原性，间接 ELISA 表明其可作为布氏杆菌病血清学诊断的候选抗原。因此，该重组蛋白是布氏杆菌病疫苗开发和血清学诊断的潜在候选抗原。
