Antibody Based Diagnosis for TB

基于抗体的结核病诊断

• 批准号: 6676191
• 负责人: Suman Laal
• 金额: $ 18.84万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6676191
• 关键词: Escherichia coli; Mycobacterium tuberculosis; antibacterial antibody; antigen antibody reaction; bacterial antigens; clinical research; diagnosis design /evaluation; diagnosis quality /standard; epitope mapping; gene expression; human tissue; molecular cloning; peptide library; proteomics; recombinant proteins; serology /serodiagnosis; synthetic peptide; tuberculosis

DESCRIPTION (provided by applicant): Attempts to devise a serodiagnostic test for TB have been made for decades with disappointing results, primarily because most of the antigens evaluated are not relevant to the human immune response. During the last 7 years, we have characterized the humoral immune responses in TB patients at different stages of active disease. These studies form the basis for the research that is now needed to develop a serodiagnostic test for TB. Our studies clearly show that antigens chosen on the basis of their immunodominance during different stages of active TB, and in both HIV-infected and non-HIV TB patients, will provide the greatest opportunity to develop a successful diagnostic test. In this context, we have already identified approximately 12 proteins in culture filtrates of M. tuberculosis that elicit antibodies in patients with incipient pre-clinical TB, non-cavitary TB or cavitary TB. This subset of antigens also elicits antibodies in patients co-infected with HIV and TB. Two proteins of this subset have been cloned, and provide a serodiagnostic assay with the highest sensitivity and specificity that has yet been achieved with any antigens. In the current application, we propose to: (a) Identify the additional immunodominant antigens. This will be done both by proteomic and molecular approaches; (b) Clone the genes for these antigens into E.coli and evaluate the reactivity of the recombinant proteins with antibodies from TB patients at different stages of TB; (c) Map the immunodominant epitopes on these candidate proteins by a variety of strategies; (d) Identify the immunodominant peptides of these antigens that are recognized by antibodies from patients across the spectrum of TB; and finally (e) Select and evaluate the combinations of peptides that will be the basis of a low-cost, rapid, point-of-care diagnostic test for TB with high sensitivity and specificity.

描述（由申请人提供）：数十年来，人们一直试图设计结核病的血清学诊断试验，但结果令人失望，主要是因为大多数评价的抗原与人体免疫应答无关。在过去的7年中，我们已经在活动性疾病的不同阶段的结核病患者的体液免疫反应的特点。这些研究构成了现在开发结核病血清学诊断试验所需的研究基础。我们的研究清楚地表明，在活动性结核病的不同阶段，以及在艾滋病毒感染和非艾滋病毒结核病患者中，基于其免疫优势选择的抗原将为开发成功的诊断测试提供最大的机会。在此背景下，我们已经在M.在早期临床前结核病、非空洞性结核病或空洞性结核病患者中引发抗体的结核病。这一抗原亚群也能激发HIV和TB共感染患者的抗体。该亚组的两种蛋白质已被克隆，并提供了具有最高灵敏度和特异性的血清学诊断测定，这是迄今为止用任何抗原实现的。在本申请中，我们提出：（a）鉴定另外的免疫显性抗原。（B）将这些抗原的基因克隆到大肠杆菌中，并评估重组蛋白与来自处于不同结核病阶段的结核病患者的抗体的反应性;（c）通过各种策略绘制这些候选蛋白上的免疫显性表位;（d）确定这些抗原中被结核病患者的抗体识别的免疫显性肽;以及最后（e）选择和评估肽的组合，所述肽的组合将是具有高灵敏度和特异性的TB的低成本、快速、即时诊断测试的基础。