An antigen-capture assay to screen donated blood for Babesia microti

用于筛查捐献血液中是否含有田鼠巴贝虫的抗原捕获测定

• 批准号: 9893818
• 负责人: Paul Michael Arnaboldi
• 金额: $ 29.98万
• 依托单位: BIOPEPTIDES, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-14 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893818
• 关键词: Adopted; Adoption; Adverse event; American; Antibodies; Antigen Targeting; Antigens; Babesia; Babesia microti; Babesiosis; Biological Assay; Blood; Blood Banks; Blood Screening; Blood Tests; Blood Transfusion; Blood donor; Borrelia microti; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chronic; Clinical; Data; Detection; Development; Disease; Doctor of Philosophy; Donor Selection; Economics; Erythrocytes; Exposure to; Geography; Goals; Human; Hybridomas; Immune; Immunological Diagnosis; Immunology procedure; Incidence; Individual; Infection; Libraries; Methods; Mus; Parasites; Patients; Performance; Phase; Procedures; Proteins; Recombinants; Red Cross; Reporter; Reporting; Research Personnel; Risk; Sampling; Sensitivity and Specificity; Serological; Severities; Ships; Specificity; Spleen; Surface; Testing; Ticks; Time; Transfusion; Vascular blood supply; antibody libraries; assay development; base; blood product; commercialization; cost; cost effective; indexing; infection risk; pathogen; performance tests; prevent; prototype; screening; tick-borne

PROJECT SUMMARY In the U.S. ~5 million individuals receive blood transfusions annually. Increasingly, these recipients are being put at risk of infection with Babesia microti, a tick-transmitted blood-borne parasite that resides in red blood cells (RBCs) and causes the disease babesiosis. B. microti represents a significant threat to the US blood supply as it survives blood banking procedures and storage conditions, and can be transmitted via transfusion. Transfusion transmitted babesiosis (TTB) is responsible for the highest percentage of transfusion-related infectious fatalities reported to the FDA. There is a critical unmet need for a method that is sensitive and specific, cost-effective, and high-throughput to screen donated blood for this pathogen. We propose the development of an EIA-based antigen-capture assay capable of detecting low levels of parasites in blood donors who are likely unaware that they are infected (e.g., asymptomatic, chronic carriers). We generated a library of antibodies using B. microti infected mice, and then screened this library to identify antibodies specific to parasite surface-expressed/secreted proteins. Using these methods, we have identified antibodies to seven B. microti proteins expressed during mammalian infection. In this study, we will generate an optimized antigen-capture assay using antibody pairs to one or more parasite secreted/surface target antigens that we have identified in preliminary studies. In Specific Aim 1, we will optimize capture and reporter antibody pairs for use in an antigen capture assay for the detection of B. microti, and confirm the specificity and sensitivity of these capture-reporter antibody pairs. In Specific Aim 2, we will performance our assay using blood obtained from patients with babesiosis, and identify a finalized antigen-capture assay to proceed to clinical assay screening with our commercialization partner in a phase II application.

项目摘要 在美国，约有500万人每年接受输血。这些收件人越来越多 面临与贝贝西亚（Babesia Microti 细胞（RBC）并引起疾病babesiosis。 B. Microti对美国的血液构成了重大威胁 供应可以在血液库的程序和存储条件下生存，并且可以通过输血来传播。 输血传播Babesiosis（TTB）负责输血相关的最高百分比 传染性死亡向FDA报告。对于一种敏感和 特定，成本效益和高通量对这种病原体捐赠的血液。我们建议 开发基于EIA的抗原捕获测定法，能够检测血液中低水平的寄生虫 可能不知道自己被感染的捐助者（例如无症状，慢性载体）。我们生成了一个 使用B. microti感染小鼠的抗体库，然后筛选该库以识别特定的抗体 表面表达/分泌的蛋白质。使用这些方法，我们已经确定了 在哺乳动物感染期间表达的七个B.微比蛋白。在这项研究中，我们将生成优化的 抗原捕获测定我们使用抗体对一个或多个寄生虫分泌/表面靶抗原的抗体对 在初步研究中已确定。在特定目标1中，我们将优化捕获和报告抗体对 用于在抗原捕获测定中用于检测微芽孢杆菌，并确认特异性和灵敏度 在这些捕获 - 重复蛋白抗体对中。在特定目标2中，我们将使用血液进行测定 从Babesiosis患者获得，并确定最终确定的抗原捕获测定法进行临床 在II期应用程序中与我们的商业化合作伙伴进行测定筛选。