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.

项目摘要/摘要 在这里，我们提出了一种快速、现场可部署、经济有效、高灵敏度和特异性的侧向流动分析方法(LFA)。 符合世界卫生组织(WHO)最近在目标产品中定义的所有要求 淋巴丝虫病(LF)监测资料(TPP)If是一个毁容和衰弱的被忽视的人 一种热带疾病，影响72个国家的4000万人，由寄生丝虫Wuchereria引起 班克罗夫蒂。为了抗击这种疾病，世卫组织在全球范围内部署了大规模药物管理(MDA)计划 规模史无前例。今天，世卫组织建议在2至10年内重复两次传播评估调查(TAS) 丙二醛停止后的3年间隔，同时承认迫切需要更多的研究来改进 TAS和治疗后监测(PTS)的诊断测试将持续到2030年及以后。2021年3月，世卫组织已 为它所寻求的测试发布了TPP，其中的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。 唯一被证明可现场部署的LF测试是免疫层析测试卡(ICT)及其后续的， 更稳定的版本是丝虫病检测试纸(FTS)。这些测试已经使用了20年来检测活跃的 感染。然而，它们不适合监测活动，因为它们只检测到这种疾病18个月。 在人被感染后，由于寄生虫负担较低，在后MDA设置中缺乏敏感度。 根据世卫组织TPP，LF监测要求RDT表明早期接触班氏假丝核菌感染性 幼虫(L3期)。目前，这种效应最好的生物标志物是针对Wb123抗原的IgG4抗体 由感染的幼虫表达。商业的IgG4/Wb123酶联免疫吸附试验存在(在Bios中)，并在该中心常规使用 用于疾病控制和预防(CDC)，但不能现场部署。一种便携的快速检测方法是由 标准诊断公司，现在是雅培公司的一部分，但没有达到预期的灵敏度，因此被放弃(SD Bioline Wb123免疫球蛋白)。 我们最近成功地开发了一种检测Wb123特异性IgG4的LFA原型，它是 其特点是分析灵敏度比雅培RDT高16-32，与商用RDT相当 Wb123酶联免疫吸附试验。我们认为这是一项重大成就。这项测试有望达到85%的临床敏感性 标准。然而，目前还不清楚它是否也会满足最低(&gt；98.8%)或理想(&gt；99.8%)的特异性标准。 我们现在提出一种双链试验，允许同时检测针对两种抗原的抗体，其中一种 将是Wb123，另一种选自一系列五种新抗原，称为WbAg1-5或WbAgX。这个 双重测试的目的是通过施加两个测试线都必须满足的条件来实现更高的特异性 为可见则将测试视为阳性，并且必须没有可见的线条才能将测试视为阴性。 我们已经获得了全球卫生特别工作组(TFGH)和美国国际开发署的拨款，用于开发一种原型 这种双链分析的版本。我们现在正在向NIH申请额外的资金，以(1)优化一个或 更多的双链分析(2)建立一个包含2000个临床样本的生物库，以证明TPP符合 所需的敏感度和特异度具有95%的统计置信度；(3)将一个双向复合体用于制造 和商业化。到第一阶段/第二阶段快速通道赠款结束时，总共将有100,000种分析可用 由TFGH、CDC和其他利益相关者进行测试，以生成保护ITS所需的现场数据 国际背书。所开发的制造工艺将能够满足预计的需求 在赞助机构可以接受的成本点上，每年进行50万-100万次分析。 值得注意的是，美国国际开发署是全球最大的低频测试客户，覆盖了全球40%以上的 需求，已经赞助了我们的初步研究，显示了他们的兴趣，并提供了一条明确的途径 商业化。简而言之，我们的RDT因此将用于监测淋巴丝虫病，这是一种 在世界卫生组织2030年被忽视的热带疾病路线图上占据突出位置，国际社会 社区在主要制药公司和非政府组织的支持下，誓言在2012年消除 伦敦宣言。