An Antigen-Detection Blood Test for Pulmonary Tuberculosis

项目摘要 我们提议的研究的目标是继续提高我们的原型的准确性和可用性 纳米盘-质谱法用于检测血液样本中的结核病。 在目标1中，我们将添加先前发现的结核分枝杆菌(Mtb)特异性胰蛋白肽 将Ag85B引入到我们的原型检测中，以提高灵敏度并探索抗原的最佳多肽数量 CFP-10、ESAT-6和Ag85B需要使用先前采集的血液来最大限度地提高检测的特异性 来自1,500名患有和不患有结核病的患者的样本(样本来自NIH资助的临床疑似 结核病研究(CSTB)R01AI053731)。在目标2中，我们将验证优化后的检测方法的性能 使用之前收集的250名患者的血液样本，并在标准情况下比较其性能 MALDI-TOF MS到Mini-12仪器(样品来自NIH资助的全球药物联盟- 耐药结核病诊断[GCDD U01AI082229])。在目标3中，我们将使用以下工具评估最终测试性能 来自摩尔多瓦、墨西哥和墨西哥诊所的3000名有结核病风险的预期登记患者的血液样本 以确定该检测方法在多大程度上区分活动性结核病患者和潜伏性结核病患者， 非结核肺部疾病和健康对照。我们的目标是提高我们的敏感性和特异性 原型化验，以满足世界卫生组织(世卫组织)结核病筛查的业绩标准 (敏感性：95%，特异性：90%)和诊断(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% ).