An Antigen-Detection Blood Test for Pulmonary Tuberculosis

项目摘要 我们提出的研究的目的是继续提高我们的原型的准确性和可用性 用于检测血液样品中的结核病（TB）的Nano-MS测定。 在目标1中，我们将添加先前鉴定的结核分枝杆菌（Mtb）特异性胰蛋白酶肽， Ag 85 B与我们的原型测定法结合，以提高灵敏度并探索抗原的最佳肽数 需要CFP-10、ESAT-6和Ag 85 B，以使用先前采集的血液最大化检测的特异性 来自1,500名结核病患者和非结核病患者的样本（来自NIH资助的临床疑似结核病患者的样本） 结核病研究（CSTB）R 01 AI 053731）。在目标2中，我们将验证优化检测试剂盒的性能 使用先前从250名患者中收集的血液样本，并将其性能与标准 MALDI-TOF MS到Mini-12仪器（来自NIH资助的全球药物联盟的样品， 耐药TB诊断[GCDD U 01 AI 082229]）。在目标3中，我们将使用以下方法评价最终测定性能： 来自摩尔多瓦、墨西哥和乌克兰诊所的3,000名前瞻性登记的结核病风险患者的血液样本， 美国以确定该检测方法如何区分活动性结核病患者和潜伏性结核病患者， 非结核性肺病和健康对照。我们的目标是提高灵敏度和特异性， 原型检测，以满足世界卫生组织（WHO）结核病筛查的性能标准 敏感性95%，特异性90%，诊断敏感性98%，特异性98%。

Project Summary The objective of our proposed study is to continue to improve the accuracy and usability of our prototype NanoDisk-MS assay for the detection of tuberculosis (TB) in blood samples. In Aim 1 we will add previously identified Mycobacterium tuberculosis (Mtb)-specific tryptic peptides of Ag85B to our prototype assay to improve sensitivity and explore the optimal number of peptides for antigens CFP-10, ESAT-6 and Ag85B required to maximize specificity of the test using previously-collected blood samples from 1,500 patients with and without TB (samples from the NIH-funded Clinical Suspicion of Tuberculosis study (CSTB) R01AI053731). In Aim 2 we will validate the performance of the optimized assay using previously collected blood samples from 250 patients and compare its performance on a standard MALDI-TOF MS to the Mini-12 instrument (samples from the NIH funded Global Consortium for Drug- resistant TB Diagnostics [GCDD U01AI082229]). In Aim 3 we will evaluate final assay performance using blood samples from 3,000 prospectively enrolled patients at risk for TB in clinics in Moldova, Mexico and the US to determine how well the assay distinguishes patients with active TB disease from those with latent TB, non-TB pulmonary diseases, and healthy controls. Our goal is to improve the sensitivity and specificity of our prototype assay in order to meet World Health Organization (WHO) performance criteria for TB screening (sensitivity:95%, specificity:90% ) and diagnosis (sensitivity:98%, specificity:98% ).