STANDARDIZED ANTIGENS FOR DIAGNOSIS OF ASPERGILLOSIS

用于诊断曲霉病的标准化抗原

• 批准号: 3134964
• 负责人: VISWANATH P KURUP
• 金额: $ 9.82万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3134964
• 关键词: Aspergillus; affinity chromatography; aspergillosis; bronchiectasis; enzyme linked immunosorbent assay; fungal antigens; immunofluorescence technique; immunoglobulin E; immunoglobulin G; immunologic skin test; monoclonal antibody; serotyping; tissue /cell culture

Aspergillus fumigatus, a ubiquitous fungus is associated with at least five different clinical forms of diseases in man, among which, allergic bronchopulmonary aspergillosis (ABPA) is the most frequently encountered. The diagnosis of ABPA depends on clinical and immunological findings. Demonstration of specific antibodies in the sera of patients is of considerable value to the diagnosis of the disease. Various antigenic preparations used in the serological tests include secretory metabolites (culture filtrate) and various extracts of the cell wall and cytoplasm. All these preparations are crude extracts and show considerable variation in their reactivity with patients' sera. These antigens also show false positive and false negative reactions in various antibody detection tests. Thus, the currently available serological tests using these antigens are neither dependable nor reliable. In order to have dependable and reliable diagnostic tests, pure and standardized antigens are necessary. The specific aims are to isolate relevant antigens and develop standardized sensitive immunological tests for diagnosis. Our approach will be to produce monoclonal antibodies (MAb) to relevant antigens of A. fumigatus utilizing hybridoma technology. At least 5 to 10 MAb against relevant antigens of A. fumigatus as determined from their reactivity with patient sera, will be produced. Using these MAb, we will isolate specific antigens through affinity chromatography. The pure antigens thus obtained will be used in immunodiagnostic methods including enzyme-linked immunosorbent assay (ELISA) and in skin testing of patients with Aspergillus-related diseases. A very sensitive biotin-avidin-linked immunosorbent assay (BALISA) will be used to detect specific IgG and IgE antibodies. With hybridomas, a continuous supply of all reagents needed for diagnostic testings can be maintained and these reagents can be easily standardized. It then will be possible to diagnose the disease more reliably by comparing the results of different laboratories. This, in turn, will help to study the well-diagnosed cases for the immunopathogenic mechanisms involved in the disease which will result in better prognosis. Our efforts will be directed toward the production of standardized reagents through the use of MAb in the diagnosis of ABPA.

烟曲霉，一种普遍存在的真菌，与至少五种 人类不同临床形式的疾病，其中，过敏 支气管肺曲霉病（ABPA）是最常见的。 ABPA的诊断依赖于临床和免疫学检查结果。 证明患者血清中存在特异性抗体， 对该病的诊断有重要价值。 各种抗原 血清学检测中使用的制剂包括分泌代谢物 （培养滤液）和细胞壁和细胞质的各种提取物。 所有这些制剂都是粗提物，并显示出相当大的差异 与患者血清的反应性。 这些抗原也显示假阳性。 各种抗体检测试验中的阳性和假阴性反应。 因此，目前可用的使用这些抗原的血清学测试是 既不可靠也不可靠。 为了有可靠和可靠的 诊断试验、纯的和标准化的抗原是必要的。 具体目标是分离相关抗原， 敏感的免疫学测试来诊断。 我们的方针是 产生针对A相关抗原的单克隆抗体（MAb）。烟曲霉 利用杂交瘤技术。 至少5至10 MAb针对相关 抗原A.根据它们与患者的反应性确定的烟曲霉菌 将生产。 利用这些单克隆抗体，我们将分离出特异性抗原 通过亲和层析。 由此获得的纯抗原将是 用于免疫诊断方法，包括酶联免疫吸附 检测（ELISA）和皮肤试验中的患者与曲霉相关 疾病 一种高灵敏度的生物素-亲和素免疫吸附试验 （BALISA）将用于检测特异性IgG和IgE抗体。 与 杂交瘤，诊断所需的所有试剂的连续供应 可以保持测试并且这些试剂可以容易地标准化。 这样就有可能通过比较 不同实验室的结果。 反过来，这将有助于研究 对于涉及免疫病理机制的确诊病例， 这种疾病将导致更好的预后。 我们的努力将是 其目的在于通过使用 单克隆抗体在ABPA诊断中的应用