M. tuberculosis exosome detection for pediatric TB diagnosis

结核分枝杆菌外泌体检测用于儿童结核病诊断

基本信息

批准号：
10640250
负责人：
Sylvia LaCourse
金额：
$ 62.91万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-21 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10640250
关键词：
5 year old Adult Bacille Calmette-Guerin vaccination Bayesian Method Biological Biological Assay Biological Markers Blood Blood specimen Cell Compartmentation Cellular Phone Cessation of life Child Childhood Classification Clinical Clinical Data Clinical Treatment Collection Cryopreservation Darkness Detection Development Diagnostic Disease Early identification Endosomes Enrollment Epidemiologic Methods Equation Evaluation Future Genetic HIV Highly Active Antiretroviral Therapy Household Immune response Immunological Models Infant Investigation Kenya Link Lipoproteins Macrophage Malignant Neoplasms Measures Methods Microscopy Modeling Molecular Monitor Morbidity - disease rate Mycobacterium tuberculosis Mycobacterium tuberculosis antigens Participant Pathogenesis Patients Pediatric cohort Performance Play Prevalence Prospective cohort Proteins Rapid diagnostics Reference Standards Research Personnel Risk Role Sampling Sensitivity and Specificity Serum Specimen Sputum Stomach Surrogate Markers Surveys Technology Testing Time Treatment Failure Tuberculosis Tuberculosis Vaccines Tuberculosis diagnosis Uncertainty Urine X Inactivation aspirate biological specimen archives biomarker discovery cohort cost diagnostic tool exosome experience extracellular vesicles field study improved lipoarabinomannan molecular diagnostics mortality mycobacterial nanotechnique novel diagnostics pediatric human immunodeficiency virus peripheral blood point of care prognostic prospective prototype respiratory response treatment response tuberculosis treatment vaccine trial

项目摘要

PROJECT SUMMARY/ABSTRACT Over one million new cases of tuberculosis (TB) and 239,000 TB-related deaths occur in children each year. Young children, especially those with HIV, are more likely to present with disseminated or extrapulmonary TB and paucibacillary disease, often missed by respiratory sampling. Non-sputum, biomarker-based diagnostic tools for rapid TB detection and treatment response in children, using easily obtained specimens are urgently needed. Exosomes are small (30-100 nm) membranous extracellular vesicles (EVs) originating from endosomal cell compartments; those secreted by M. tuberculosis (Mtb) or Mtb-infected macrophages appear to play a significant role in Mtb pathogenesis. Our collaborators have developed a rapid and sensitive nanoplasmon-enhanced scattering (nPES) assay which directly detects Mtb-exosomes (Mtb-EVs) from as little as 1 L of serum. Proof- of-concept nPES assays performed with Mtb markers LpqH (19-kDa Mtb lipoprotein) and LAM distinguished adult TB from at-risk patients and normal controls, and among pediatric TB cases (including HIV+) and controls with high sensitivity and specificity. We propose using archived specimens and clinical data from the Pediatric Urgent Start of HAART (PUSH) Study (NCT02063880) and a new proposed prospective cohort of children suspected of TB with high HIV prevalence to evaluate performance of nPES detected Mtb-EVs for pediatric TB diagnosis (Aim 1), treatment response (Aim 2), and evaluation of a point-of-care platform (Aim 3). In addition to assessing conventional diagnostic performance measures, we propose to use advanced epidemiologic methods (Bayesian latent class analysis) given the context of an imperfect reference. Additional evaluation in adult TB patients and healthy controls including household contacts (adults and children) and recently BCG-vaccinated infants without TB is proposed. We hypothesize nPES detected Mtb-EVs will 1) have similar diagnostic performance to the reference of Xpert/culture among children with confirmed TB without the need for sputum, and identify additional children missed by respiratory sample, 2) will provide a useful surrogate marker of treatment response with decline in quantitative levels during successful treatment, and 3) will maintain performance with a point-of-care platform. Using cryopreserved samples from a well-characterized cohort of children with HIV who underwent intensive TB evaluation and a prospective cohort of children suspected of TB with high HIV prevalence provides opportunity for efficient evaluation of a novel diagnostic with potential for clinical impact to improve pediatric TB diagnosis.

项目摘要/摘要每年儿童发生超过100万例新的结核病（TB）和239,000个与TB有关的死亡。幼儿，尤其是艾滋病毒的孩子，更有可能出现散布或肺外结核和paucibaCillarry疾病，通常被呼吸抽样遗漏。非估计，基于生物标志物的诊断工具对于儿童的快速TB检测和治疗反应，迫切需要使用轻松获得的标本。外泌体是源自内体细胞的小（30-100 nm）膜细胞外蔬菜（EV）车厢；那些由结核分枝杆菌（MTB）或MTB感染的巨噬细胞分泌的人似乎起着重要的作用在MTB发病机理中的作用。我们的合作者开发了一种快速而敏感的纳米素增强散射（NPE）测定，直接从低至1 l的血清中检测到MTB-exosomes（MTB-EV）。证明- 用MTB标记LPQH（19-KDA MTB脂蛋白）和LAM区分的概念NPESSSEASS 来自风险患者和正常对照组以及小儿结核病病例（包括HIV+）和对照组的成人结核具有高灵敏度和特异性。我们建议使用HAART小儿紧急开始的存档标本和临床数据（PUSH）研究（NCT02063880）和涉嫌高HIV患病率的TB的新的拟议的前瞻性队列评估NPE检测到的MTB-EV的性能用于小儿结核病诊断（AIM 1），治疗反应（AIM 2），以及评估即时平台（AIM 3）。除了评估常规诊断绩效指标，我们建议使用高级流行病学方法（贝叶斯潜在类别分析）给定不完美的参考的上下文。成人结核病患者和健康对照组的其他评估提出了包括家庭接触（成人和儿童）和最近没有结核病的BCG接种疫苗的婴儿。我们假设检测到的NPE的MTB-EVS 1）具有与参考的诊断性能相似确认结核病儿童中的XPERT/文化无需痰液，并确定其他儿童被呼吸样本错过，2）将提供有用的治疗反应的替代标记，而下降成功治疗期间的定量水平和3）将通过护理点平台保持性能。使用来自艾滋病毒的儿童的冷冻保存样本，这些样本接受了密集的结核病评估和涉嫌高HIV患病率的涉嫌结核病的儿童提供了机会为了有效评估具有临床影响的新型诊断，以改善小儿结核病诊断。