Real-time Detection of Active TB in HIV Exposed Children on Customized Nanotrap

使用定制 Nanotrap 实时检测 HIV 暴露儿童的活动性结核病

• 批准号: 8846187
• 负责人: Tony Y. Hu
• 金额: $ 39.88万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-09 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8846187
• 关键词: Abate; Address; Adult; Affect; Age; Algorithms; Aliquot; Antibodies; Antigens; Antitubercular Agents; Applications Grants; Automobile Driving; Bacteria; Bacterial DNA; Base Sequence; Biological Assay; Biological Markers; Biology; Biopsy; Blinded; Blood; Blood Circulation; Blood specimen; Characteristics; Child; Childhood; Clinical; Clinical Chemistry; Clinical Management; Communicable Diseases; Communities; Couples; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Disease; Drug resistance in tuberculosis; Early identification; Early treatment; Evaluation; Exhibits; Extreme drug resistant tuberculosis; Genus Mycobacterium; Goals; Gold; HIV; Health; Healthcare; Human; Individual; Infection; International; Knowledge; Label; Life; Light; Logistic Regressions; Lung; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Measures; Methods; Modality; Monitor; Multidrug-Resistant Tuberculosis; Mycobacterium tuberculosis; Nanotechnology; Nature; Patients; Peptides; Perinatal; Plasma; Prevention; Probability; Procedures; Process; Property; Protocols documentation; Recombinants; Records; Regimen; Relative (related person); Risk; Risk Assessment; Risk Factors; Sampling; Sensitivity and Specificity; Serum; Signal Transduction; Silicon Dioxide; Specificity; Specimen; Spinal Puncture; Stomach; Symptoms; Taxes; Techniques; Technology; Testing; Therapeutic; Thoracic Radiography; Time; Translations; Treatment Efficacy; Tuberculin Test; Tuberculosis; Validation; Variant; Virus Diseases; Work; base; care burden; clinical Diagnosis; clinically relevant; co-infection; cohort; global health; improved; insight; interest; light curve; lymph nodes; multiplex detection; nanopore; patient population; pediatric patients; prevent; research study; respiratory; response; sample collection; screening; tool; transmission process; tuberculosis treatment; validation studies

DESCRIPTION (provided by applicant): Tuberculosis (TB), particularly multidrug and extensively drug-resistant TB (M/XDR-TB), poses a significant, worldwide health threat to HIV-exposed children, as progression from infection to disease can occur rapidly and unpredictably. The current gold standard for TB diagnosis and treatment monitoring in children as well as adults relies heavily on time-consuming bacterial culturing or bacterial DNA detection (e.g. GeneXpert MTB/RIF) methods. Clinical diagnosis of childhood TB is especially challenging because of the paucibacillary (few bacteria) nature of the disease and difficulties in obtaining clinically relevant specimens. Even sample collection is itself an enormous task because it often requires invasive procedures (e.g., gastric aspiration, lymph node biopsy, and lumbar puncture). To minimize the destruction and health care burdens resulting from this infectious disease in vulnerable HIV-infected children, a specific and quantitative diagnostic test for active TB is needed for early identification and prevention of TB transmission. We have recently developed a biomarkers detection assay that couples the tunable properties of silica nanopore chips (referred to as Nanotrap ) to the detection capabilities of our benchtop matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS), which capitalizes on recent advances in clinical chemistry applied to MS that enables a different biomarkers detection modality not seen to date. Our preliminary studies reveal three major findings: (1) with the Nanotrap-MS platform, two TB biomarkers (CFP-10 and ESAT-6) can be detected in circulation at an incredibly low concentration (as little as 1.0 fmol), flagging the presence of possible infection well before any conventional technique can assess bacterial load; (2) we have used the platform to differentiate 51 HIV+/TB+ children from 27 HIV+/TB- children and 18 healthy controls simply by detecting the presence of CFP-10 and ESAT-6 in patients' blood, rather than performing burdensome bacterial isolation procedures, with 100% specificity and 94% sensitivity; and (3) perhaps most strikingly, the time between sample processing to answer, or diagnosis, is only half a day rather than the typical 4-6 week period required for conventional methods! In this proposal, we aim to advance the Nanotrap-MS platform much closer to full clinical translation by: a) establishing a simple but extremely accurate protocol to quantify the amount of CFP-10 and ESAT-6 in tandem in a small blood aliquot; b) performing a clinical validation study for Nanotrap- MS, using a large sample set collected from TB-infected children who may also be HIV carriers; and c) investigating the platform's feasibility in monitoring the (early) treatment efficacy of anti-TB therapies, as well as screening for the possible emergence of M/XDR-TB. Successful execution of the proposed studies will help to diminish the likelihood of or even prevent TB transmission, and improve clinical management of TB. Furthermore, the Nanotrap-MS platform has enormous potential not just for TB disease assessment, but it also has broad applications in virtually any clinical indication that manifests in biomarker release int circulation.

描述（由申请人提供）：结核病（TB），特别是耐多药和广泛耐药结核病（M/XDR-TB），对艾滋病毒暴露儿童构成重大的全球健康威胁，因为从感染到疾病的进展可能会迅速发生和不可预测的。目前儿童和成人结核病诊断和治疗监测的黄金标准在很大程度上依赖于耗时的细菌培养或细菌 DNA 检测（例如 GeneXpert MTB/RIF）方法。儿童结核病的临床诊断尤其具有挑战性，因为该疾病具有少杆菌（少数细菌）性质并且难以获得临床相关标本。即使是样本采集本身也是一项艰巨的任务，因为它通常需要侵入性操作（例如胃抽吸、淋巴结活检和腰椎穿刺）。为了最大限度地减少这种传染病对易受艾滋病毒感染的儿童造成的破坏和医疗负担，需要对活动性结核病进行特异性和定量的诊断测试，以便及早识别和预防结核病传播。我们最近开发了一种生物标志物检测方法，将二氧化硅纳米孔芯片（称为 Nanotrap）的可调特性与我们的台式基质辅助激光解吸/电离飞行时间质谱 (MALDI-TOF MS) 的检测能力结合起来，它利用了应用于 MS 的临床化学的最新进展，实现了迄今为止未见过的不同生物标志物检测方式。我们的初步研究揭示了三个主要发现：(1) 利用 Nanotrap-MS 平台，可以在循环中以极低的浓度（低至 1.0 fmol）检测到两种结核病生物标志物（CFP-10 和 ESAT-6），从而标记在任何常规技术可以评估细菌负荷之前就已经存在可能的感染； (2) 我们使用该平台，只需检测患者血液中 CFP-10 和 ESAT-6 的存在，即可将 51 名 HIV+/TB+ 儿童与 27 名 HIV+/TB- 儿童和 18 名健康对照区分开来，而不是进行繁琐的细菌分离程序具有 100% 的特异性和 94% 的敏感性； (3) 也许最引人注目的是，从样本处理到回答或诊断之间的时间仅为半天，而不是传统方法所需的典型 4-6 周时间！在本提案中，我们的目标是通过以下方式使 Nanotrap-MS 平台更接近完全临床转化： a) 建立一个简单但极其准确的方案来量化小份血液中 CFP-10 和 ESAT-6 的含量； b) 使用从结核感染儿童（也可能是 HIV 携带者）收集的大量样本对 Nanotrap-MS 进行临床验证研究； c) 调查该平台在监测抗结核疗法（早期）疗效以及筛查可能出现的耐多药/广泛耐药结核方面的可行性。成功执行拟议研究将有助于减少甚至预防结核病传播的可能性，并改善结核病的临床管理。此外，Nanotrap-MS 平台不仅在结核病评估方面具有巨大潜力，而且在循环中生物标志物释放的几乎所有临床适应症中也具有广泛的应用。