Ultrasensitive, High-Specificity Rapid Test to Support the End-Game of the Global Program to Eliminate Lymphatic Filariasis

超灵敏、高特异性快速检测，支持全球消除淋巴丝虫病计划的最后阶段

• 批准号: 10480110
• 负责人: Marco Antonio Biamonte
• 金额: $ 25.96万
• 依托单位: BIG EYE DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10480110
• 关键词: Achievement; Address; Advisory Committees; Affect; Aftercare; Antibodies; Antigens; Award; Benchmarking; Biological Assay; Biological Markers; Centers for Disease Control and Prevention (U.S.); Clinical; Clinical Sensitivity; Collaborations; Communities; Country; Data; Detection; Devices; Diagnostic; Diagnostic tests; Disease; Ensure; Enzyme-Linked Immunosorbent Assay; Exposure to; Filaria bancrofti; Filarial Elephantiases; Filariasis; Freezing; Funding; Grant; Human; IgG4; Immunoglobulin G; Infection; International; Investments; Larva; Life; London; Low Prevalence; Medical Device; National Institute of Allergy and Infectious Disease; Parasites; Performance; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Procedures; Proteins; Publishing; Rapid diagnostics; Recrudescences; Research; Sampling; Secure; Sensitivity and Specificity; Series; Specificity; Speed; Stress Tests; Surveys; Testing; Time; Tropical Climate; USAID; United States National Institutes of Health; Work; World Health Organization; antibody detection; base; biobank; combat; commercialization; cost; cost effective; design; detection assay; disease diagnostic; falls; global health; improved; interest; lateral flow assay; manufacturing process; neglected tropical diseases; novel; novel diagnostics; pathogen; portability; programs; prototype; rapid test; repository; response; test strip; transmission process; volunteer

PROJECT SUMMARY / ABSTRACT Here we propose a rapid, field-deployable, cost-effective, highly sensitive and specific lateral flow assay (LFA) that meets all the requirements recently defined by the World Health Organization (WHO) in a target product profile (TPP) for surveillance activities of Lymphatic Filariasis (LF). LF is a disfiguring and debilitating neglected tropical disease that affects 40 million people in 72 countries and is due to the parasitic filarial worm Wuchereria bancrofti. To combat this disease, WHO has deployed mass drug administration (MDA) programs globally on an unprecedented scale. Today, WHO recommends repeating transmission assessment surveys (TAS) twice in 2- to 3-year intervals after MDA cessation, while acknowledging that more research is urgently required to improve diagnostic tests for TAS and post-treatment surveillance (PTS) until 2030, and beyond. In March 2021, WHO has published a TPP for the test it seeks, where the key requirements are that the test should be able to detect early infection, have >85% clinical sensitivity and > 98.8% specificity. The only proven field-deployable tests for LF are the immunochromatography testcard (ICT) and its subsequent, more stable version, the Filariasis Test Strip (FTS). These tests have been used for 2 decades to detect active infections. However, they are not suited for surveillance activities, as they detect the disease only 18 months after a person has been infected and lack sensitivity in post-MDA settings due to low parasite burden. According to the WHO TPP, LF surveillance requires an RDT indicative of early exposure to W. bancrofti infective larvae (L3 stage). Currently, the best biomarker to this effect are IgG4 antibodies specific for the Wb123 antigen expressed by infective larvae. A commercial IgG4/Wb123 ELISA exists (inBios) and is routinely used at the Centers for Disease Control and Prevention (CDC) but is not field-deployable. A portable rapid test was introduced by Standard Diagnostics, now part of Abbott, but fell short of the expected sensitivity and was abandoned (SD BIOLINE Wb123 IgG). We have recently succeeded in developing an LFA prototype for the detection of Wb123-specific IgG4 which is characterized by an analytical sensitivity 16-32 greater than that of the Abbott RDT and equal to the commercial Wb123 ELISA. We view this as a major achievement. This test is poised to meet the > 85% clinical sensitivity criterion. However, it is not clear if it will also meet the minimal (>98.8%) or ideal (>99.8%) specificity criterion. We now propose a biplex test allowing to simultaneously detect antibodies against two antigens, of which one will be Wb123, and the other one chosen from a series of five novel antigens, termed WbAg1-5 or WbAgX. The purpose of the dual test is to achieve a superior specificity by imposing the condition that both test lines must be visible to count the test as positive, and no line must be visible for the test to be counted as negative. We have been awarded a grant from the Task Force for Global Health (TFGH) and USAID to develop a prototype version of such a biplex assay. We are now requesting additional funding from the NIH to (1) optimize one or more biplex assays (2) establish a biorepository of 2000 clinical samples to demonstrate that the TPP meets the required sensitivity and specificity with > 95% statistical confidence and (3) progress one biplex to manufacturing and commercialization. By the end of this Phase I/II Fast-Track grant, a total of 100,000 assays will be available for testing by the TFGH, the CDC, and other stakeholders to generate the field data necessary to secure its international endorsement. The developed manufacturing process will be able to meet the estimated demand of 0.5-1 million assays/year at a cost point acceptable to the sponsoring agencies. It is noteworthy that USAID, which is the largest customer for LF tests worldwide, covering over 40% of the global demand, has already sponsored our initial research, showing their interest, and providing a clear path to commercialization. In short, our RDT will thus be poised for the surveillance of lymphatic filariasis, a disease that figures prominently on the WHO’s 2030 roadmap for Neglected Tropical Diseases, and which the international community, supported by major pharmaceutical companies and NGOs, has vowed to eliminate in the 2012 London Declaration.

项目概要/摘要 在这里，我们提出了一种快速、可现场部署、经济高效、高灵敏度和特异性的侧流检测 (LFA) 满足世界卫生组织 (WHO) 最近在目标产品中定义的所有要求 淋巴丝虫病（LF）监测活动的概况（TPP）是一个被忽视的毁容和衰弱的疾病。 影响 72 个国家 4000 万人的热带疾病，由寄生丝虫 Wuchereria 引起 为了对抗这种疾病，世卫组织在全球范围内部署了大规模药物管理（MDA）计划。 如今，世卫组织建议每 2 至 2 次重复进行传播评估调查 (TAS) 两次。 停止 MDA 后间隔 3 年，同时认识到迫切需要更多研究来改进 世卫组织已在 2030 年及以后进行 TAS 诊断检测和治疗后监测 (PTS)。 为其寻求的测试发布了 TPP，其中关键要求是测试应该能够及早发现 感染，具有 >85% 的临床敏感性和 > 98.8% 的特异性。 唯一经过验证的可现场部署的 LF 测试是免疫层析测试卡 (ICT) 及其后续产品， 更稳定的版本，丝虫病测试条 (FTS) 这些测试已使用了 20 年来检测活动性丝虫病。 然而，它们不适合监测活动，因为它们仅在 18 个月内发现疾病。 一个人被感染后，由于寄生虫负担低，在 MDA 后的环境中缺乏敏感性。 根据 WHO TPP，LF 监测需要进行 RDT，以表明早期接触过班克罗夫氏菌传染性 目前，达到此效果的最佳生物标志物是针对 Wb123 抗原的 IgG4 抗体。 存在由感染性幼虫表达的商业 IgG4/Wb123 ELISA (inBios)，并且在中心常规使用。 用于疾病控制和预防 (CDC)，但不可现场部署 便携式快速测试由 标准诊断公司 (Standard Diagnostics)，现已成为雅培 (Abbott) 的一部分，但由于未达到预期的灵敏度而被放弃 (SD BIOLINE Wb123 IgG）。 我们最近成功开发了用于检测 Wb123 特异性 IgG4 的 LFA 原型，该原型是 其特点是分析灵敏度比 Abbott RDT 高 16-32，等于商业产品 我们认为这是一项重大成就，该测试有望满足 > 85% 的临床敏感性。 然而，尚不清楚它是否也满足最低（> 98.8％）或理想（> 99.8％）特异性标准。 我们现在提出了一种双联测试，可以同时检测针对两种抗原的抗体，其中一种是 一个是 Wb123，另一个是选自一系列五种新抗原，称为 WbAg1-5 或 WbAgX。 双重测试的目的是通过施加两条测试线必须的条件来实现卓越的特异性 必须可见才能将测试计为阳性，并且必须没有可见线才能将测试计为阴性。 我们获得了全球健康工作组 (TFGH) 和美国国际开发署的资助，用于开发原型 我们现在请求 NIH 提供额外资金以 (1) 优化一种或多种方法。 更多双重分析 (2) 建立 2000 个临床样本的生物样本库，以证明 TPP 符合 所需的灵敏度和特异性具有 > 95% 的统计置信度，并且 (3) 将一种双链体推进到生产 到第一阶段/第二阶段快速通道资助结束时，总共将有 100,000 项检测可用。 供 TFGH、CDC 和其他利益相关者进行测试，以生成确保其安全所需的现场数据 所开发的制造工艺将能够满足预计的需求。 每年进行 0.5-100 万次检测，成本点可以为主办机构所接受。 值得注意的是，USAID是全球LF测试最大的客户，覆盖全球40%以上 需求，已经赞助了我们的初步研究，表现出了他们的兴趣，并提供了明确的途径 简而言之，我们的 RDT 将用于监测淋巴丝虫病，这种疾病 世界卫生组织 2030 年被忽视的热带病路线图占据显着位置，国际社会 在主要制药公司和非政府组织的支持下，社区发誓要在 2012 年消除 伦敦宣言。