Peptide-Based Serodiagnostic Test for Cysticercosis

描述（由申请人提供）：该项目的目的是利用合成和重组蛋白作为抗原开发新的标准化猪囊尾蚴病血清学检测方法。将从对应于CDC蛋白质印迹的七个诊断条带的组分中选择抗原，所述CDC蛋白质印迹利用从来源于受感染猪的寄生虫分离的小扁豆凝集素糖蛋白-目前用于检测针对T的幼虫阶段的抗体的最灵敏和特异性的血清学试验。索利亚将开发两种类型的检测：标准微孔板ELISA形式和膜试纸条检测。创新的免疫捕获检测技术将应用于新的检测，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方案，扩大使用纯合成和重组抗原的优势。快速简便的ELISA检测将理想地满足血清流行病学调查和社区研究期间对多个样本进行实验室检测以及诊断囊虫感染的要求。试纸测试将提供一个方便和更具成本效益的替代目前的蛋白质印迹测试，并将适用于小批量或个别测试在实验室或现场条件。将在新检测中实施的抗体检测原则将提供对人类和动物样本使用相同检测的可能性。该项目的第1阶段包括分析合成和重组蛋白抗原、鉴定较短的免疫显性序列、合成ELISA和膜基检测试剂，以及选择适用于检测人和猪样本的抗原和检测配置的最佳组合。第2阶段将包括对选定格式的额外优化、使用更大的人类和猪样本组进行验证以及在现场条件下进行临床评价。

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.