Peptide-Based Serodiagnostic Test for Cysticercosis

描述(由申请人提供)：该项目的目标是利用合成和重组蛋白作为抗原，开发新的囊虫病标准化血清学检测方法。抗原将从CDC Western Blot的七条诊断条带对应的成分中选择，该方法利用从受感染猪的寄生虫中分离出的小扁豆凝集素糖蛋白-目前最敏感和特异的血清学检测方法，用于检测针对猪弓形虫幼虫阶段的抗体。将开发两种类型的检测：标准微孔板ELISA格式和膜试纸试验。一种创新的免疫捕获检测技术将被应用于新的检测，通过进一步减少通常伴随血清学检测的非特异性反应，并通过显著简化检测程序，放大使用纯合成和重组抗原的优势。在血清流行病学调查和基于社区的研究中，这种快速和简单的ELISA检测将理想地满足对多个样本进行实验室测试的要求，以及诊断囊虫病感染。试纸测试将为目前的Western Blot测试提供一种方便和更具成本效益的替代方法，并适用于实验室或现场条件下的小批量或个别测试。将在新的检测中实施的抗体检测原理将使对人类和动物样本使用相同的检测成为可能。该项目的第一阶段包括分析合成和重组蛋白抗原，鉴定较短的免疫优势序列，合成用于ELISA和基于膜的检测的试剂，以及选择适用于检测人和猪样本的最佳抗原组合和测试配置。第二阶段将包括进一步优化选定的格式，使用更大的人和猪样本小组进行验证，以及在现场条件下进行临床评估。

DESCRIPTION (provided by applicant): The aim of this project will be development of new standardized serological tests for cysticercosis utilizing synthetic and recombinant proteins as antigens. Antigens will be selected from components corresponding to seven diagnostic bands of the CDC Western Blot, which utilizes lentil lectin glycoproteins isolated from parasites derived from infected pigs - currently the most sensitive and specific serological test for detection of antibodies against the larval stage of T. solium. Two types of tests will be developed: a standard microplate ELISA format, and a membrane dipstick test. An innovative immune capture assay technology will be applied for new tests which will amplify the advantages of using pure synthetic and recombinant antigens by further reduction of the non-specific reactions which usually accompany serological tests, and by significant simplification of assay protocols. The quick and simple ELISA test will ideally meet requirements for laboratory testing of multiple samples during seroepidemiological surveys and community-based studies, as well as for diagnosis of cysticercosis infection. The dipstick test will provide a convenient and more cost-effective alternative to the current Western Blot test, and will be suitable for small batch or individual testing in lab or field conditions. Principles of antibody detection which will be implemented in the new tests will provide the possibility to use the same tests on both human and animal samples. Phase 1 of this project includes analysis of synthetic and recombinant protein antigens, identification of shorter immunodominant sequences, synthesis of reagents for ELISA and membrane-based tests, and selection of optimal combinations of antigens and test configurations applicable for testing human and porcine samples. Phase 2 will comprise additional optimization of selected formats, validation using larger panels of human and porcine specimens, and clinical evaluation under field conditions.