Antibody Based Diagnosis for TB

基于抗体的结核病诊断

• 批准号: 6855759
• 负责人: Suman Laal
• 金额: $ 45.12万
• 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/6855759
• 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）：几十年来，人们一直在尝试设计结核病血清诊断测试，但结果令人失望，主要是因为大多数评估的抗原与人类免疫反应无关。在过去的 7 年里，我们描述了结核病患者在活动性疾病不同阶段的体液免疫反应。这些研究构成了现在开发结核病血清诊断检测所需的研究基础。我们的研究清楚地表明，在活动性结核病的不同阶段以及艾滋病毒感染者和非艾滋病毒结核病患者中，根据其免疫优势选择抗原将为开发成功的诊断测试提供最大的机会。在这种情况下，我们已经在结核分枝杆菌培养滤液中鉴定出大约 12 种蛋白质，这些蛋白质可在早期临床前结核病、非空洞结核病或空洞结核病患者中引发抗体。这部分抗原也会在同时感染艾滋病毒和结核病的患者中引发抗体。该子集的两种蛋白质已被克隆，并提供了迄今为止任何抗原都达到的最高灵敏度和特异性的血清诊断测定。在当前的应用中，我们建议：（a）识别额外的免疫显性抗原。这将通过蛋白质组学和分子方法来完成； (b) 将这些抗原的基因克隆到大肠杆菌中，并评估重组蛋白与结核病不同阶段的结核病患者的抗体的反应性； (c) 通过多种策略绘制这些候选蛋白上的免疫显性表位； (d) 鉴定这些抗原的免疫显性肽，这些肽可被所有结核病患者的抗体识别；最后 (e) 选择并评估肽的组合，这些肽的组合将成为低成本、快速、高灵敏度和特异性的结核病护理点诊断测试的基础。