First-in-class, highly sensitive and specific rapid diagnostic test to detect Paragonimus infections

用于检测并殖吸虫感染的一流、高灵敏度和特异性的快速诊断测试

基本信息

批准号：
10545410
负责人：
Marco Antonio Biamonte
金额：
$ 25.82万
依托单位：
BIG EYE DIAGNOSTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-14 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545410
关键词：
Address Adult Affect Antibiotics Antibodies Antigens Appearance Area Asia Biological Assay Biological Markers Blood Body Weight decreased Caspase Centers for Disease Control and Prevention (U.S.)Chemotherapy-Oncologic Procedure Clinical Clinical Management Clinical Sensitivity Collaborations Collection Coughing Country Crustacea Cyst Data Detection Development Diagnosis Diagnostic Diagnostic tests Differential Diagnosis Exposure to Feces Filarial Elephantiases Fresh Water Funding Goals Gold Hand Hemoptysis High Prevalence Human IgG4 Impairment Infection Ingestion Intervention Intestinal Volvulus Lung Lung diseases Malignant neoplasm of lung Medical Methodology Microscopy Myoglobin Nematode infections Onchocerciasis Optics Paragonimiasis Paragonimus Patients Performance Persons Pharmaceutical Preparations Phase Philippines Praziquantel Prevalence Proteins Public Health Quality of Care Rapid diagnostics Reader Regimen Reporter Research Resources Route Sampling Sensitivity and Specificity Serology Serum Southeastern Asia Specificity Sputum Symptoms Technology Testing Trematoda Tuberculosis Tuberculosis diagnosis Universities Vendor Washington Work X-Ray Computed Tomography assay development base biobank college commercialization cost effective density detection method egg experience fighting genome sequencing global health immunoreactivity improved infection rate interest lateral flow assay medical schools nanoparticle nanoshell neglect neglected tropical diseases novel plasmonics point of care professor programs prototype undercooked volunteer

项目摘要

PROJECT SUMMARY / ABSTRACT Herein, we propose an ultrasensitive serological rapid diagnostic test (RDT) to detect exposure to pulmonary flukes of the genus Paragonimus. Paragonimiasis affects an estimated 21 million people and remains amongst the most neglected of all neglected tropical diseases (NTDs). Trematodes of the genus Paragonimus are acquired upon ingestion of raw or undercooked freshwater crustaceans and migrate to the lungs where they encyst. Treatment is straightforward (praziquantel) but diagnosis is not. The classical method – detection of Paragonimus eggs in sputum or stool – requires rarely available microscopy expertise, is slow and hence low-throughput. With an effective drug at hand but no convenient diagnostic, a highly sensitive and specific RDT is the missing piece to enable efficient clinical management of paragonimiasis. RDTs represent a mature, robust, and cost-effective technology ideal for intervention at the point of care in resource-limited countries. An additional important problem is that many paragonimiasis symptoms resemble those of TB or lung cancer, including hemoptysis (blood in sputum), persistent cough, weight loss, and cysts in CT scans. As a result, misdiagnosis is common and patients are incorrectly put on long and ineffective antibiotic regimens or on toxic cancer chemotherapy, with abysmal consequences for the patients. The importance of differential diagnosis between paragonimiasis and other pulmonary diseases further underscores the need for a Paragonimus RDT. The project will leverage novel paragonimiasis biomarkers, notably a cysteine protease (CP-6) and myoglobin-1, recently discovered at the Washington University School of Medicine after an extensive effort of genome sequencing. In previous work, we developed an RDT prototype that detects IgG4 antibodies against CP-6 with 89% sensitivity and 100% specificity when evaluated on a small panel of positive (n=19) and negative (n=28) sera. The RDT utilizes a novel lateral-flow assay (LFA) set-up combined with novel plasmonic reporter nanoparticles (gold nanoshells) for superior optical detection. However, some test lines were weak, and further optimization is required to warrant manufacturing. Even more importantly, assay development was severely limited by the paucity of clinical samples. Therefore, a particular focus of this proposal is to assemble a collection of paragonimiasis sera, in collaboration with Dr. Belizario, Dean and Professor, College of Public Health, University of the Philippines, Manila. We are therefore requesting funding to assemble such a biorepository of both paragonimiasis-positive and -negative patient samples (Specific Aim 1), to further optimize the sensitivity of the prototype RDT without impairing the specificity, e.g., by generating a biplex assay to simultaneously detect IgG4 antibodies against both antigens, CP-6 (test line 1) and myoglobin-1 (test line 2), whereby the appearance of either test line would be counted as positive (Specific Aim 2), and to validate and compare the different assays generated in Aim 2 using the samples acquired under Aim 1, and select the best prototype for further development in a subsequent Phase II (Specific Aim 3). Dr. Belizario indicates that for a Paragonimus RDT to have value in the field, it should have a sensitivity of minimally >80% and ideally >90%, and a specificity of >97%. A simple test for paragonimiasis remains an unmet medical need, and by addressing this need, we hope to improve the quality of care of people with pulmonary diseases in parts of Southeast Asia. Although high prevalence areas in Asia stand to benefit most from a new test, the assay would also be useful in the USA, despite low infection rates, as witnessed by the CDC’s interest in this project. Furthermore, although the WHO has not yet officially endorsed paragonimiasis as an NTD requiring a global surveillance and/or elimination program, our RDT is nevertheless prone to be of interest to the WHO in the context of their massive global effort to fight TB, as it will allow a better differential diagnosis of TB vs. Paragonimiasis cases. Our team has the track record, the technological expertise and the global health experience required to deliver a successful product.

项目概要/摘要在此，我们提出了一种超灵敏的血清学快速诊断测试（RDT）来检测肺吸虫的暴露情况并殖吸虫属感染估计有 2100 万人，并且仍然是最容易被忽视的疾病之一。所有被忽视的热带病（NTD）都是通过摄入生的或吸虫属的吸虫而感染的。未煮熟的淡水甲壳类动物并迁移到肺部，在那里形成包囊。治疗很简单。（吡喹酮），但诊断不需要经典方法——检测痰或粪便中的并殖吸虫卵。很少有可用的显微镜专业知识，速度慢，因此通量低。手头上有有效的药物，但没有方便的诊断方法，高度敏感和特异性的 RDT 是 RDT 代表了一种成熟、稳健且具有成本效益的技术，能够对肺吸虫病进行有效的临床管理。是在资源有限的国家进行护理点干预的理想选择。肺吸虫病的症状与结核病或肺癌相似，包括咯血（痰中带血）、持续咳嗽、体重减轻和 CT 扫描中出现囊肿，因此误诊很常见，并且患者会被错误地长时间佩戴。无效的抗生素疗法或有毒的癌症化疗，给患者带来可怕的后果。肺吸虫病和其他肺部疾病之间鉴别诊断的重要性进一步强调了需要对于并殖吸虫 RDT。该项目将利用新型并殖吸虫病生物标志物，特别是半胱氨酸蛋白酶 (CP-6) 和肌红蛋白-1，最近华盛顿大学医学院经过大量的基因组测序工作后发现了这一点。在这项工作中，我们开发了一种 RDT 原型，可以检测 CP-6 的 IgG4 抗体，灵敏度为 89%，特异性为 100% 当对一小部分阳性 (n=19) 和阴性 (n=28) 血清进行评估时，RDT 采用了一种新型侧流检测。（LFA）装置与新型等离子体报告纳米颗粒（金纳米壳）相结合，可实现卓越的光学检测。然而，一些测试线很薄弱，需要进一步优化才能保证生产。重要的是，临床样本的缺乏严重限制了测定的发展，因此，特别关注。该提案旨在与院长兼教授 Belizario 博士合作收集肺吸虫病血清，菲律宾大学公共卫生学院，马尼拉。因此，我们请求资金来组装这样一个兼具阳性和阴性肺吸虫病的生物储存库患者样本（具体目标 1），以进一步优化原型 RDT 的灵敏度而不损害特异性，例如，通过生成双重检测来同时检测针对两种抗原的 IgG4 抗体，CP-6（测试线 1）和肌红蛋白-1（测试线 2），其中任何一条测试线的出现都将被视为阳性（具体目标 2），并且使用目标 1 下采集的样本验证和比较目标 2 中生成的不同测定，并选择 Belizario 博士指出，这是后续第二阶段进一步开发的最佳原型（具体目标 3）。并殖吸虫 RDT 要在现场具有价值，其灵敏度应至少 >80%，理想情况下 >90%，并且特异性 >97%。肺吸虫病的简单检测仍然是一个未满足的医疗需求，通过解决这一需求，我们希望改善东南亚部分地区肺部疾病患者的护理质量，尽管亚洲的患病率较高的地区仍然如此。为了从新测试中获益最多，该检测方法在美国也很有用，尽管感染率较低，正如此外，尽管世界卫生组织尚未正式认可肺吸虫病作为一种疾病，但疾病预防控制中心对此项目很感兴趣。 NTD 需要全球监测和/或消除计划，但我们的 RDT 很容易引起人们的兴趣世卫组织在其全球抗击结核病的大规模努力的背景下，因为这将有助于更好地鉴别诊断结核病和结核病。我们的团队拥有所需的记录、技术专长和全球健康经验。交付成功的产品。