Validation of urine/serum LAM in HIV/nonHIV TB suspects and POC Test Development

HIV/非 HIV 结核病疑似者的尿液/血清 LAM 验证和 POC 测试开发

基本信息

批准号：
10179309
负责人：
DELPHI CHATTERJEE
金额：
$ 55.87万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-08 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179309
关键词：
Acid Fast Bacillae Staining Method Address Adult Affinity Antibodies Area Bacillus Bacteria Bacterial DNA Binding Biological Assay Biological Markers Blood Cell Wall Chemistry Child Clinical Collaborations Complex Country DNA Decontamination Detection Detergents Development Devices Diagnosis Diagnostic Diagnostic tests Disease Drug resistance in tuberculosis Engineering Enzyme-Linked Immunosorbent Assay Epidemic Epitopes Evaluation Goals Gold Growth HIV HIV Seronegativity HIV Seropositivity HIV/TB Immune response Kenya Knowledge Mass Fragmentography Memory B-Lymphocyte Methodology Methods Microbiology Microscopy Molecular Cloning Monoclonal Antibodies Mycobacterium tuberculosis Nature Patients Peptide Hydrolases Peru Population Procedures Production Property Protein Denaturation Proteins Protocols documentation Reagent Reference Standards Reporting Research Sampling Sensitivity and Specificity Serum South Africa Specificity Specimen Sputum Testing Thailand Tuberculosis Urea Urine Validation Work antibody detection base cell envelope clinical Diagnosis clinical diagnostics cohort cost design detection limit diagnostic accuracy diagnostic assay diagnostic platform drug development extensive drug resistance human subject immunosuppressed improved in vivo interest lateral flow assay lipoarabinomannan pandemic disease point of care point of care testing prevent protein complex resistant strain treatment response tuberculosis diagnostics tuberculosis drugs tuberculosis treatment urinary

项目摘要

Despite the emergence of multiple and extensively drug resistant strains, tuberculosis (TB) is largely a curable disease if treated appropriately. However, in some tuberculosis-endemic areas, fewer than 40% of TB cases are diagnosed due to the lack of accurate and easy-to-use diagnostic assays, and these continue to drive the epidemic. Currently, diagnosis relies on demonstration of the bacteria, Mycobacterium tuberculosis, in clinical specimens by sputum smear microscopy and repeat culture. Limitations of culture often include slow growth (up to 8 weeks), low sensitivity as a result of required decontamination procedures, difficulty in obtaining serial samples in children and non-sputum producers, and the high cost of the assay. A non-sputum, non-growth based biomarker assay could replace microbiologic intermediate endpoints, could transform the pace and scope of TB drug development and of global TB control. It may additionally have utility for reaching a population of paucibacillary disease as is often seen in children, extrapulmonary TB, and HIV/TB. This proposed research will establish our goal towards developing a highly sensitive approach to the quantitative detection of lipoarabinomannan (LAM), a cell envelope lipoglycan in serum and urine with clinical diagnostic accuracy. The project aims are 1) to optimize the urine and serum pretreatment protocol by testing proteases and chaotropic agents, thus making maximal amount of the LAM available for detection; 2) to generate highly specific and unique monoclonal antibodies directed against “in vivo” forms of LAM; and 3) to take forward non-sputum based LAM detection via a capture ELISA with further validation in well-characterized adult cohorts with suspected TB in countries which have high burdens of TB and HIV (South Africa, Thailand, Kenya) and (Peru) with low HIV exposure. Given the recognized imperfection of culture as a gold standard, in a separate exploratory analysis we will determine ELISA sensitivity and specificity using LAM detection by Gas Chromatography/Mass Spectrometry (GC/MS) as the reference standard. Our capture ELISA accuracy targets are ≥85% sensitivity and ≥95% specificity. In addition, we will determine, the changes in urine and serum LAM concentrations during TB treatment; this will use a separate set of existing specimens and is intended to establish whether LAM in urine might be useful as a biomarker of treatment response. Our ultimate goal is to develop an accurate, simple Point-of-Care test based on quantitative engineering of capture chemistry and conditions for LAM detection in non-sputum specimens. The test should reach all TB cases irrespective of their HIV status. The chosen demonstration will not be a final device but will demonstrate a proof-of-principle that is aligned with the design goals.

尽管出现了多种广泛耐药菌株，但结核病 (TB) 在很大程度上是可治愈的然而，在一些结核病流行地区，结核病病例的比例不到 40%。由于缺乏准确且易于使用的诊断方法而被诊断出来，这些继续推动检测目前，诊断依赖于临床上结核分枝杆菌的证明。通过痰涂片显微镜检查和重复培养来获取标本，培养的局限性通常包括生长缓慢。（长达 8 周），由于所需的净化程序而导致灵敏度低，难以获得序列号儿童样本无痰，且检测成本高，无痰，不生长。基于生物标志物测定可以取代微生物学中间终点，可以改变速度和它可能还有助于实现结核病药物开发和全球结核病控制的范围。少杆菌病人群（常见于儿童、肺外结核病和艾滋病毒/结核病）。这项拟议的研究将确立我们的目标，即开发一种高度敏感的方法来应对定量检测脂阿拉伯甘露聚糖包膜（LAM），一种血清和尿液中的细胞脂聚糖，具有临床意义该项目的目标是 1) 通过测试优化尿液和血清预处理方案。蛋白酶和离液剂，从而使最大量的 LAM 可用于检测；产生针对“体内”形式的 LAM 的高度特异性和独特的单克隆抗体；以及 3) 通过捕获 ELISA 推进非基于痰的 LAM 检测，并在充分表征的情况下进行进一步验证结核病和艾滋病毒高负担国家（南非、泰国、肯尼亚）和（秘鲁）的艾滋病毒暴露率较低，鉴于文化作为黄金标准的缺陷已得到公认。在单独的探索性分析中，我们将使用气体 LAM 检测来确定 ELISA 的敏感性和特异性色谱/质谱 (GC/MS) 作为参考标准我们的捕获 ELISA 准确性目标。敏感性≥85%，特异性≥95% 此外，我们将确定尿液和血清 LAM 的变化。结核病治疗期间的浓度；这将使用一组单独的现有样本，旨在确定尿液中的 LAM 是否可用作治疗反应的生物标志物。基于捕获化学的定量工程开发准确、简单的即时检测非痰标本中 LAM 检测的条件该检测应覆盖所有结核病病例，无论其年龄如何。所选择的演示不是最终设备，而是演示原理验证。与设计目标保持一致。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Estimation of D-Arabinose by Gas Chromatography/Mass Spectrometry as Surrogate for Mycobacterial Lipoarabinomannan in Human Urine.

通过气相色谱/质谱法估算 D-阿拉伯糖作为人尿液中分枝杆菌脂阿拉伯甘露聚糖的替代物。

DOI：
发表时间：
2015
期刊：
影响因子：
3.7
作者：
De, Prithwiraj;Amin, Anita G;Valli, Eloise;Perkins, Mark D;McNeil, Michael;Chatterjee, Delphi
通讯作者：
Chatterjee, Delphi

Urine lipoarabinomannan as a marker for low-risk of NTM infection in the CF airway.

尿液脂阿拉伯甘露聚糖作为 CF 气道 NTM 感染低风险的标志物。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
De, Prithwiraj;Amin, Anita G;Graham, Barbara;Martiniano, Stacey L;Caceres, Silvia M;Poch, Katie R;Jones, Marion C;Saavedra, Milene T;Malcolm, Kenneth C;Nick, Jerry A;Chatterjee, Delphi
通讯作者：
Chatterjee, Delphi

Urine lipoarabinomannan in HIV uninfected, smear negative, symptomatic TB patients: effective sample pretreatment for a sensitive immunoassay and mass spectrometry.

未感染 HIV、涂片阴性、有症状结核病患者的尿液脂阿拉伯甘露聚糖：用于灵敏免疫测定和质谱分析的有效样品预处理。

DOI：
发表时间：
2021-02-03
期刊：
影响因子：
4.6
作者：
Amin, Anita G;De, Prithwiraj;Graham, Barbara;Calderon, Roger I;Franke, Molly F;Chatterjee, Delphi
通讯作者：
Chatterjee, Delphi

Simple manipulation of enzyme-linked immunosorbent assay (ELISA) using an automated microfluidic interface.

使用自动化微流体接口对酶联免疫吸附测定 (ELISA) 进行简单操作。

DOI：
发表时间：
2022-05-13
期刊：
影响因子：
0
作者：
Panraksa, Yosita;Jang, Ilhoon;Carrell, Cody S;Amin, Anita G;Chailapakul, Orawon;Chatterjee, Delphi;Henry, Charles S
通讯作者：
Henry, Charles S

Comparative Structural Study of Terminal Ends of Lipoarabinomannan from Mice Infected Lung Tissues and Urine of a Tuberculosis Positive Patient.

来自感染小鼠肺组织和结核阳性患者尿液的脂阿拉伯甘露聚糖末端的比较结构研究。