STANDARDIZED ANTIGENS FOR DIAGNOSIS OF ASPERGILLOSIS

烟曲霉，一种普遍存在的真菌，与至少五种 人类疾病的不同临床形式，其中，过敏性 最常见的是支气管肺曲霉菌病(ABPA)。 ABPA的诊断依赖于临床和免疫学结果。 患者血清中的特异性抗体的证明是 对该病的诊断有相当大的价值。各种抗原性 血清学检测中使用的制剂包括分泌型代谢物 (培养滤液)以及细胞壁和细胞质的各种提取物。 所有这些制剂都是粗提物，表现出相当大的差异。 它们与病人血清的反应性。这些抗原也显示出假的 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。为了有靠得住、靠得住 诊断性试验、纯净和标准化的抗原是必要的。 其具体目的是分离相关抗原，并建立标准化 敏感的免疫学诊断试验。我们的方法将是 产生抗烟曲霉菌相关抗原的单抗 利用杂交瘤技术。至少5到10个MAB，与相关 从烟曲霉菌与病人的反应性确定其抗原 血清，将被生产出来。利用这些单抗，我们将分离出特定的抗原。 通过亲和层析。这样获得的纯抗原将是 用于包括酶联免疫吸附在内的免疫诊断方法 曲霉菌相关性患者的检测(ELISA)和皮肤试验 疾病。一种非常灵敏的生物素-亲和素偶联免疫吸附试验 (BALISA)将用于检测特定的免疫球蛋白抗体和免疫球蛋白E抗体。使用 杂交瘤，持续供应诊断所需的所有试剂 检测可以维护，这些试剂可以很容易地标准化。 然后，通过比较，可以更可靠地诊断这种疾病 不同实验室的结果。这反过来又将有助于研究 被充分诊断的病例涉及免疫致病机制 这种疾病会导致更好的预后。我们的努力将是 用于生产标准化试剂通过使用 单抗在ABPA诊断中的价值。

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.