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

• 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
• 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

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.

项目摘要/摘要 在此，我们提出了一种超灵敏的血清学快速诊断试验(RDT)来检测肺吸虫暴露。 并殖吸虫属。据估计，肺吸虫病影响着2100万人，并且仍然是最被忽视的 所有被忽视的热带疾病(NTDS)。并殖吸虫属的吸虫是通过摄入生食或 未煮熟的淡水甲壳类动物并迁移到肺部，在那里它们被包囊。治疗方法简单明了 (吡喹酮)但诊断并非如此。经典的方法--检测痰或粪便中的并殖吸虫卵--需要 几乎没有可用的显微镜专业知识，速度很慢，因此吞吐量低。 由于手头有有效的药物，但没有方便的诊断，高度敏感和特异的RDT是缺少的一块 实现对并殖吸虫病的有效临床管理。RDT代表了一种成熟、健壮且经济高效的技术 非常适合在资源有限的国家进行护理时进行干预。另外一个重要的问题是，许多 肺吸虫病的症状类似于结核病或肺癌，包括咯血(痰中带血)、持续咳嗽、 体重减轻，还有CT扫描中的囊肿症。因此，误诊很常见，患者被错误地放在长时间和 无效的抗生素方案或有毒的癌症化疗，给患者带来可怕的后果。这个 肺吸虫病与其他肺部疾病的鉴别诊断的重要性进一步强调了 对于一只并殖吸虫RDT。 该项目将利用新的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 在华盛顿大学医学院经过广泛的基因组测序工作后发现的。在以前的 在这项工作中，我们开发了一种RDT原型，它检测CP-6的IgG4抗体具有89%的敏感性和100%的特异性 在一小群阳性(n=19)和阴性(n=28)血清上进行评估时。RDT采用了一种新的侧向流动分析方法 (LFA)设置与新型等离子体报告纳米颗粒(金纳米壳)相结合，实现卓越的光学检测。 然而，一些测试线很弱，需要进一步优化才能保证生产。甚至更多 重要的是，临床样本的缺乏严重限制了化验技术的发展。因此，一个特别的焦点是 这项提议是与院长兼教授贝里萨里奥博士合作，收集并殖吸虫病血清， 菲律宾大学公共卫生学院，马尼拉。 因此，我们正在申请资金，以建立这样一个肺吸虫病阳性和阴性并殖吸虫病的生物库。 患者样本(特定目标1)，为了在不损害特异性的情况下进一步优化原型RDT的灵敏度， 例如，通过产生同时检测针对CP-6(测试线1)和Cp-6两种抗原的IgG4抗体的双链试验 肌红蛋白-1(测试线2)，因此任何一条测试线的出现将被视为阳性(特定目标2)，以及 使用在目标1下采集的样本来验证和比较在目标2中生成的不同化验结果，并选择 在随后的第二阶段进一步开发的最佳原型(具体目标3)。贝里萨里奥博士指出，对于一个 RDT并殖吸虫要在现场有价值，它的敏感度最低应为80%，理想情况下应为90%，并且 &gt；的特异性为97%。 对并殖吸虫病的简单测试仍然是一个未得到满足的医疗需求，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。尽管亚洲的高发地区 为了最大限度地受益于一项新的检测，该检测方法在美国也将是有用的，尽管感染率很低，正如 疾控中心对这个项目的兴趣。此外，尽管世卫组织尚未正式认可并殖吸虫病为 NTD需要全球监测和/或消除计划，我们的RDT仍然容易引起 世卫组织在其抗击结核病的大规模全球努力的背景下，因为这将使对结核病的鉴别诊断与 肺吸虫病病例。我们的团队拥有所需的记录、技术专长和全球卫生经验 交付一款成功的产品。