Mycobacterium bovis (M. bovis), the main cause of animal tuberculosis (TB), can infect a wide variety of domestic and wild animal species, including suids. Suids may serve as reservoir hosts or disease sentinels in different scenarios. Accurate detection of M. bovis infection in pigs is important for TB control programs. Although previous studies have shown the value of serological assays for screening animal populations, the diagnostic accuracy was considered suboptimal. In this study, we used Dual Path Platform (DPP) technology and multi-antigen print immunoassay (MAPIA) to characterize antigen recognition profiles and temporal antibody responses. Four M. bovis experimentally infected pigs developed an early antibody response to antigen MPB83, with a peak in IgG levels starting around 4–6 weeks post-inoculation, although none of the pigs developed antibodies to fusion protein CFP10/ESAT6 within 16 weeks of the experiment. Three of four experimentally infected pigs developed antibody responses before detectable antigen-specific interferon gamma responses. Naturally infected pigs with gross lesions containing viable M. bovis showed IgM (19/40 infected animals) and IgG (39/40) antibody responses to both MPB70/MPB83 (39/40) and CFP10/ESAT6 (34/40). Using MPB70/MPB83 antigen alone to measure IgG antibody levels by DPP assay, an estimated test sensitivity was 97.5 % (95 % CI: 85.3−99.9 %). None of the 57 negative control samples had detectable IgM or IgG antibodies to either of the two test antigens in DPP assay, suggesting an estimated specificity of 100 % (95 % CI: 92.1–100.0 %) in pigs. MAPIA showed robust IgG reactivity to multiple protein antigens of M. bovis in the naturally infected pigs. The results demonstrate that serological assays which detect IgG antibodies to MPB83 have high sensitivity and specificity for accurate detection of M. bovis infection in pigs. Further investigations should be done to validate anti-MPB70/MPB83 antibodies as a reliable serodiagnostic biomarker for TB diagnosis in pigs.

牛分枝杆菌（牛分枝杆菌）是动物结核病（TB）的主要原因，可感染包括suid在内的多种家养和野生动物物种。在不同情况下，水体可能充当水库宿主或疾病哨兵。准确检测猪的牛分枝杆菌感染对于结核病控制计划很重要。尽管先前的研究表明血清学检测对筛查动物种群的价值，但诊断准确性被认为不是最佳的。在这项研究中，我们使用了双通道平台（DPP）技术和多抗原印刷免疫分析（MAPIA）来表征抗原识别谱和瞬时抗体反应。四牛经实验感染的猪对MPB83抗原产生早期抗体反应，IgG水平在接种后约4-6周开始达到峰值，尽管在实验的16周内没有猪对融合蛋白CFP10 / ESAT6产生抗体。在实验感染的四只猪中，有三只在可检测到的抗原特异性干扰素γ反应之前就已经产生了抗体反应。自然感染的猪，其病灶中含有活牛分枝杆菌展示了对MPB70 / MPB83（39/40）和CFP10 / ESAT6（34/40）的IgM（感染19/40只动物）和IgG（39/40）抗体的反应。仅使用MPB70 / MPB83抗原通过DPP测定来测量IgG抗体水平，估计的测试灵敏度为97.5％（95％CI：85.3-99.9％）。在DPP分析中，这57个阴性对照样品均未检测到针对两种测试抗原的IgM或IgG抗体，这表明在猪中的特异性估计为100％（95％CI：92.1-100.0％）。MAPIA在天然感染的猪中显示出对牛分枝杆菌的多种蛋白抗原具有很强的IgG反应性。结果表明，检测针对MPB83的IgG抗体的血清学检测方法对准确检测牛分枝杆菌具有很高的灵敏度和特异性。猪感染。应该做进一步的研究，以验证抗MPB70 / MPB83抗体是猪结核病诊断的可靠血清诊断生物标志物。