Background Biomarker-based tests for diagnosing TB currently rely on detecting Mycobacterium tuberculosis (Mtb) antigen-specific cellular responses. While this approach can detect Mtb infection, it is not efficient in diagnosing TB, especially for patients who lack aetiological evidence of the disease. Methods We prospectively enrolled three cohorts for our study for a total of 630 subjects, including 160 individuals to screen protein biomarkers of TB, 368 individuals to establish and test the predictive model and 102 individuals for biomarker validation. Whole blood cultures were stimulated with pooled Mtb-peptides or mitogen, and 640 proteins within the culture supernatant were analysed simultaneously using an antibody-based array. Sixteen candidate biomarkers of TB identified during screening were then developed into a custom multiplexed antibody array for biomarker validation. Results A two-round screening strategy identified eight-protein biomarkers of TB: I-TAC, I-309, MIG, Granulysin, FAP, MEP1B, Furin and LYVE-1. The sensitivity and specificity of the eight-protein biosignature in diagnosing TB were determined for the training (n=276), test (n=92) and prediction (n=102) cohorts. The training cohort had a 100% specificity (95% CI 98% to 100%) and 100% sensitivity (95% CI 96% to 100%) using a random forest algorithm approach by cross-validation. In the test cohort, the specificity and sensitivity were 83% (95% CI 71% to 91%) and 76% (95% CI 56% to 90%), respectively. In the prediction cohort, the specificity was 84% (95% CI 74% to 92%) and the sensitivity was 75% (95% CI 57% to 89%). Conclusions An eight-protein biosignature to diagnose TB in a high-burden TB clinical setting was identified.

中文翻译：

---

鉴定用于诊断结核病的八种蛋白质生物印记

背景 用于诊断结核病的基于生物标志物的测试目前依赖于检测结核分枝杆菌 (Mtb) 抗原特异性细胞反应。虽然这种方法可以检测 Mtb 感染，但它在诊断 TB 方面效率不高，尤其是对于缺乏该疾病病因学证据的患者。方法 我们前瞻性地为我们的研究招募了三个队列，共 630 名受试者，其中 160 名用于筛选 TB 的蛋白质生物标志物，368 名用于建立和测试预测模型，102 名用于生物标志物验证。用混合的 Mtb 肽或有丝分裂原刺激全血培养物，并使用基于抗体的阵列同时分析培养物上清液中的 640 种蛋白质。然后将筛选期间确定的 16 种候选结核病生物标志物开发成定制的多重抗体阵列，用于生物标志物验证。结果 两轮筛选策略确定了 TB 的八种蛋白质生物标志物：I-TAC、I-309、MIG、Granulysin、FAP、MEP1B、Furin 和 LYVE-1。针对训练 (n=276)、测试 (n=92) 和预测 (n=102) 队列确定了八种蛋白质生物印记在诊断 TB 中的敏感性和特异性。使用随机森林算法方法通过交叉验证，训练队列具有 100% 的特异性（95% CI 98% 至 100%）和 100% 敏感性（95% CI 96% 至 100%）。在测试队列中，特异性和敏感性分别为 83%（95% CI 71% 至 91%）和 76%（95% CI 56% 至 90%）。在预测队列中，特异性为 84%（95% CI 74% 至 92%），敏感性为 75%（95% CI 57% 至 89%）。结论 鉴定了在高负担结核病临床环境中诊断结核病的八种蛋白质生物印记。