Peptide-Based Serodiagnostic Test for Cysticercosis

基于肽的囊尾蚴病血清诊断测试

• 批准号: 6933533
• 负责人: Victor Andrew KOvalenko
• 金额: $ 13.62万
• 依托单位: IMMUNETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933533
• 关键词: Cestoda; clinical research; diagnosis design /evaluation; diagnostic tests; enzyme linked immunosorbent assay; helminth genetics; helminthic antigen; human tissue; recombinant proteins; synthetic peptide; western blottings

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.

描述（由申请人提供）：本项目旨在利用合成蛋白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7个诊断条带对应的成分中选择，该方法利用从感染猪的寄生虫中分离的扁豆凝集素糖蛋白，这是目前用于检测针对血吸虫幼虫期抗体的最敏感和特异性的血清学检测方法。将开发两种类型的测试：标准的微孔板ELISA格式和膜试纸测试。一种创新的免疫捕获测定技术将应用于新的测试，通过进一步减少通常伴随血清学测试的非特异性反应，并通过大大简化分析方案，将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的要求，以及用于囊虫病感染的诊断。试纸测试将为目前的Western Blot测试提供一种方便且更具成本效益的替代方法，并且适用于实验室或现场条件下的小批量或单个测试。将在新测试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测试的可能性。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ELISA和膜检测试剂，选择适用于检测人和猪样品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，使用更大的人类和猪样本进行验证，以及在现场条件下进行临床评估。