Bovine tuberculosis (bTB) control programs can be improved by implementation of advanced ante-mortem testing algorithms. Serodiagnostic methods using traditional blood or blood-derived specimens may benefit from the use of less invasive alternative biological fluids, provided those mirror systemic antibody responses. In the present study, we used Dual Path Platform (DPP) and Multiantigen Print Immunoassay (MAPIA) to compare antibody levels in ten sample types including whole blood (fresh and hemolyzed), plasma (fresh and leftover from Bovigam testing), serum, saliva, broncho-alveolar lavage, urine, diaphragm extract, and bile collected from cattle aerosol-infected with Mycobacterium bovis. High correlation (r = 0.97−0.99) in measurements of IgG antibodies to MPB70/MPB83 fusion antigen by DPP assay was found between all blood-derived specimens, supporting matrix equivalency. Broncho-alveolar lavage and diaphragm extract yielded positive results in all the infected animals tested, showing high correlation with matching serum data (r = 0.94 and r = 0.95, respectively) and suggesting their potential use in antibody assays. Characterized by MAPIA, the antigen reactivity patterns obtained with paired sera and alternative specimens were nearly identical, with slight differences in intensity. Antibodies were also found by DPP assay in saliva, urine, and bile from some of the infected animals, but the titers were relatively low, thus reducing the diagnostic value of such specimens. The proposed approach was evaluated in a pilot field study on warthogs diagnosed with M. bovis infection. Relative levels of antibody in tissue fluid obtained from lymph nodes or lungs were consistent with those detected in sera and detectable in all infected warthogs. The findings support the diagnostic utility of non-traditional biological fluids and tissue samples when used as alternative test specimens in serologic assays for bTB.

牛结核病 (bTB) 控制程序可以通过实施先进的宰前测试算法来改进。使用传统血液或血液衍生样本的血清诊断方法可能会受益于使用侵入性较小的替代生物液体，前提是那些反映全身抗体反应。在本研究中，我们使用双通道平台 (DPP) 和多抗原印迹免疫分析 (MAPIA) 来比较十种样本类型中的抗体水平，包括全血（新鲜和溶血）、血浆（新鲜和来自 Bovigam 检测的剩余）、血清、唾液、支气管肺泡灌洗液、尿液、隔膜提取物和从牛分枝杆菌气溶胶感染的牛收集的胆汁. 通过 DPP 测定法测量 MPB70/MPB83 融合抗原的 IgG 抗体的高相关性（r = 0.97-0.99）在所有血液来源的样本之间发现，支持基质等效性。支气管肺泡灌洗液和隔膜提取物在所有测试的感染动物中均产生了阳性结果，显示出与匹配的血清数据高度相关（分别为 r = 0.94 和 r = 0.95），并表明它们在抗体测定中的潜在用途。以 MAPIA 为特征，用配对血清和替代标本获得的抗原反应模式几乎相同，强度略有不同。部分感染动物的唾液、尿液和胆汁中也发现了抗体，但滴度较低，降低了此类标本的诊断价值。牛分枝杆菌感染。从淋巴结或肺获得的组织液中抗体的相对水平与在血清中检测到的一致，并且在所有感染的疣猪中都可以检测到。这些发现支持非传统生物体液和组织样本在 bTB 血清学检测中用作替代测试样本时的诊断效用。