Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.
J Clin Microbiol. 2019 Sep 24;57(10). doi: 10.1128/JCM.00274-19. Print 2019 Oct.
Bovine tuberculosis (bTB) is caused by During the early stage of infection, greater than 15% of -infected cattle shed mycobacteria through nasal secretions, which can be detected by nested PCR. To compare the differences in the protein profiles of -infected cattle that were nested PCR positive (bTB) and -infected cattle that were nested PCR negative (bTB) and to screen for biomarkers that will facilitate the early and accurate detection of bTB, we investigated the protein expression profiles of serum and bovine purified protein derivative (PPD-B)-stimulated plasma among bTB ( = 20), bTB ( = 20), and uninfected cattle (NC; = 20) by iTRAQ labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2D LC-MS/MS). After comprehensive analysis, we selected 15 putative differentially expressed serum proteins and 15 plasma proteins for validation by parallel reaction monitoring (PRM) with the same cohort used in the iTRAQ analysis. Four serum and five PPD-B-stimulated proteins were confirmed in follow-up enzyme-linked immunosorbent assays. PPD-B-stimulated interleukin 8 (IL-8) displayed the potential to differentiate -infected cattle from NC, with an area under the curve (AUC) value of 0.9662, while PPD-B-stimulated C-reactive protein (CRP) displayed the potential to differentiate bTB from bTB, with an AUC value of 1.00. Finally, double-blind testing with 244 cattle indicated that the PPD-B-stimulated IL-8 test exhibited good agreement with traditional tests (κ > 0.877) with a >90% relative sensitivity and a >98% relative specificity; the PPD-B-stimulated CRP test displayed good agreement with nested PCR (κ = 0.9117), with an observed 94% relative sensitivity and 97% relative specificity. Therefore, the PPD-B-stimulated IL-8 and CRP tests could be used to detect bTB and to differentiate bTB from bTB.
牛结核病（bTB）是由引起的。在感染的早期阶段，超过 15%的感染牛通过鼻腔分泌物排出分枝杆菌，这可以通过巢式 PCR 检测到。为了比较巢式 PCR 阳性（bTB）和巢式 PCR 阴性（bTB）感染牛之间的蛋白质谱差异，并筛选有助于早期准确检测 bTB 的生物标志物，我们通过 iTRAQ 标记结合二维液相色谱-串联质谱法（iTRAQ-2D LC-MS/MS）研究了 bTB（n=20）、bTB（n=20）和未感染牛（NC；n=20）的血清和牛纯化蛋白衍生物（PPD-B）刺激的血浆中的蛋白质表达谱。经过综合分析，我们选择了 15 种可能差异表达的血清蛋白和 15 种血浆蛋白，并用相同的 iTRAQ 分析队列进行平行反应监测（PRM）验证。在后续的酶联免疫吸附试验中，验证了 4 种血清蛋白和 5 种 PPD-B 刺激蛋白。PPD-B 刺激白细胞介素 8（IL-8）显示出区分感染牛和 NC 的潜力，曲线下面积（AUC）值为 0.9662，而 PPD-B 刺激 C 反应蛋白（CRP）显示出区分 bTB 和 bTB 的潜力，AUC 值为 1.00。最后，对 244 头牛进行的双盲测试表明，PPD-B 刺激的 IL-8 测试与传统测试具有良好的一致性（κ>0.877），相对敏感性>90%，相对特异性>98%；PPD-B 刺激的 CRP 测试与巢式 PCR 具有良好的一致性（κ=0.9117），观察到的相对敏感性为 94%，相对特异性为 97%。因此，PPD-B 刺激的 IL-8 和 CRP 测试可用于检测 bTB，并区分 bTB 和 bTB。
