Peptide-Based Serological Test for Cysticercosis

基于肽的囊尾蚴病血清学检测

• 批准号: 7274759
• 负责人: Victor Andrew KOvalenko
• 金额: $ 32.81万
• 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 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7274759
• 关键词: Agreement; Antibodies; Antigen-Antibody Complex; Antigens; Area; Attention; Biological Assay; Blood Tests; Calcified; Centers for Disease Control and Prevention (U.S.); Central Nervous System Infections; Cerebrospinal Fluid; Clinical; Clinical Research; Clinical Trials; Collaborations; Community Surveys; Condition; Cyst; Cysticercosis; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic Reagent; Diagnostic tests; Differential Diagnosis; Discrimination; Disease; Echinococcosis; Enzyme-Linked Immunosorbent Assay; Epidemiologic Studies; Equipment; Family suidae; Future; Goals; Health; Helminths; Human; Human Resources; Immune; In Vitro; Individual; Infection; International; Label; Laboratories; Larva; Lateral; Licensing; Membrane; Neurocysticercosis; Neurologic; Neuroschistosomiasis; Numbers; Pan American Health Organization; Parasites; Parasitic Diseases; Patients; Peptides; Performance; Phase; Preparation; Protocols documentation; Purpose; Reaction; Reagent; Recombinant Proteins; Recombinants; Research; Research Personnel; Sampling; Sensitivity and Specificity; Serodiagnoses; Serologic tests; Serum; Severities; Specificity; Specimen; Staging; Standardization; Standards of Weights and Measures; Sus scrofa; Synthetic Antigens; Taenia solium; Techniques; Technology; Testing; Training; United States Food and Drug Administration; Western Blotting; Whole Blood; World Health Organization; base; commercialization; cost; day; human disease; innovation; manufacturing process; synthetic peptide; technology validation

DESCRIPTION (provided by applicant): The pork tapeworm Taenia solium is now increasingly recognized as a parasite with international health implications and the capability of spreading to non-endemic areas. The growing attention to this helminth species relates primarily to the severity of health problems associated with infection of the central nervous system with the larval stage, which causes neurocysticercosis currently classified as the most important neurologic disease of parasitic origin. In spite of the theoretical possibility for eradication of T. solium cysticercosis global eradication of this infection is still unlikely in the near future. The overall goal of the current Phase II application will be to finish the development, validate through clinical trials, and launch the manufacture of serological tests for detection of specific antibodies to circulating antigens of the larval stage of Taenia solium. The tests will utilize standardized antigenic material such as synthetic peptides, recombinant proteins and combinations thereof selected from antigens identified recently in diagnostic bands of Western blots (WB) prepared from cyst material of naturally infected pigs. Two types of tests will be developed: one will be in a standard microwell ELISA format, and the second will be a rapid non-instrumental format utilizing lateral flow principles. An innovative but already proven immune-capture assay technology will be applied in both new tests, which will emphasize the advantage of using pure synthetic and recombinant antigens. The quick and simple ELISA test will ideally meet the requirements for laboratory testing of multiple samples in sero-epidemiological surveys and community-based studies, as well as for diagnosis of cysticercosis infection in individuals. The lateral flow format with capability for whole blood testing will provide a convenient and more cost-effective alternative to the current WB test, and will be suitable for small batch or individual testing in labs or field conditions. Methodological innovations that will be implemented in the lateral flow test will make its sensitivity equal to or even higher than the sensitivity of the microplate ELISA version. Both ELISA and lateral flow tests will be able to detect antibodies in samples from two hosts of T. solium, humans and pigs. Phase II of this project includes finalizing the analysis of synthetic and recombinant antigens in order to select combination(s) with a minimal number of antigens providing equal or better sensitivity than the current WB test for detection of antibodies in patients with various stages of disease, with comparably high specificity. The second major goal will be validation of technology for preparation of antigenic material and diagnostic reagents including launching the manufacture of pilot lots of the new tests. Significant efforts at this stage will be directed towards optimization and manufacture of highly sensitive lateral flow versions of the serological tests. The objective of the present project is the development of the two types of a commercial serological tests for diagnosis of infection caused by larva stage of pork tapeworm Taenia solium in human and pigs utilizing standardized antigenic material such as synthetic peptides and recombinant proteins. The first test will be in simple microwell ELISA format well suited for testing of any number of samples, using semi-automated or automated equipment in clinical labs and equipped facilities. The second test will be in rapid non-instrumental format utilizing lateral flow principles for application in field conditions or in laboratories for testing relatively small batches of samples. The new tests will replace the expensive WB technique that requires specially trained personnel, and will make possible a level of standardization and accuracy in clinical and reference laboratories that is currently unattainable. The primary application of these tests will be the diagnosis of neurocysticercosis in humans, pig cysticercosis, seroepidemiological surveys and community-based studies.

描述（由申请人提供）：猪绦虫现在越来越多地被认为是一种具有国际卫生影响的寄生虫，并且能够传播到非流行地区。越来越多的关注，这种蠕虫物种主要涉及到严重的健康问题与感染的中枢神经系统与幼虫阶段，这导致神经囊尾蚴病目前被列为最重要的神经系统疾病的寄生虫来源。尽管理论上存在根除T.猪囊尾蚴病在全球范围内根除这种感染在不久的将来仍然是不可能的。目前第二阶段申请的总体目标是完成开发，通过临床试验进行验证，并开始生产血清学检测试剂，用于检测猪带绦虫幼虫期循环抗原的特异性抗体。该测试将利用标准化的抗原材料，例如合成肽、重组蛋白及其组合，其选自最近在由自然感染的猪的包囊材料制备的蛋白质印迹（WB）的诊断条带中鉴定的抗原。将开发两种类型的测试：一种是标准微孔ELISA格式，第二种是利用侧向流原理的快速非仪器格式。一种创新的但已经被证明的免疫捕获检测技术将被应用于这两种新的检测中，这将强调使用纯合成和重组抗原的优势。这种快速简便的ELISA检测方法将理想地满足血清流行病学调查和社区研究中对多个样本进行实验室检测的要求，以及个人囊虫病感染的诊断要求。具有全血检测能力的侧流格式将为当前WB检测提供方便且更具成本效益的替代方案，并且将适用于实验室或现场条件下的小批量或个体检测。将在侧向流测试中实施的方法学创新将使其灵敏度等于或甚至高于微孔板ELISA版本的灵敏度。ELISA和侧流试验都能够检测来自两种宿主T.人和猪的关系该项目的第二阶段包括完成合成和重组抗原的分析，以选择具有最少数量抗原的组合，提供与当前WB检测相同或更好的灵敏度，用于检测不同疾病阶段患者的抗体，具有相对较高的特异性。第二个主要目标是验证抗原材料和诊断试剂的制备技术，包括启动新检测试剂的试验批次的生产。在这一阶段的重大努力将致力于优化和生产高灵敏度的侧流版本的血清学测试。本项目的目的是利用标准化抗原材料，如合成肽和重组蛋白，开发两种类型的商业血清学试验，用于诊断人和猪中由猪绦虫幼虫期引起的感染。第一次检测将采用简单的微孔ELISA形式，非常适合在临床实验室和装备设施中使用半自动或自动设备检测任何数量的样本。第二次测试将采用快速非仪器形式，利用横向流动原理，在现场条件下或在实验室中测试相对较小批次的样品。新的测试将取代昂贵的WB技术，需要经过专门培训的人员，并将使临床和参考实验室的标准化和准确性水平成为可能，这是目前无法实现的。这些检测的主要应用将是人类脑囊虫病、猪囊虫病的诊断、血清流行病学调查和社区研究。