Point-of-sampling detection of Lyme and other tick-borne diseases

莱姆病和其他蜱传疾病的采样点检测

• 批准号: 9763438
• 负责人: Ozlem Yaren
• 金额: $ 18.5万
• 依托单位: FOUNDATION FOR APPLIED MOLECULAR EVOLUTN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-14 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763438
• 关键词: Advocacy; American; Ammonia; Anaplasma; Antibodies; Architecture; Awareness; Babesia; Babesiosis; Bacteria; Benchmarking; Biohazardous Substance; Biological Assay; Blood; Borrelia; Borrelia burgdorferi; Borrelia miyamotoi; Buffers; Centers for Disease Control and Prevention (U.S.); Child; China; Communicable Diseases; Computer software; Culicidae; Data; Dengue; Detection; Devices; Diagnosis; Diagnostic; Disease; Drops; Environment; Ethanol; FDA approved; Florida; Fluorescence; Foundations; Freeze Drying; Health; Heating; Home environment; Human; India; Information Systems; Laboratories; Left; Lyme Disease; Molecular; Molecular Evolution; Monitor; National Institute of Allergy and Infectious Disease; Nurses; One-Step dentin bonding system; Oranges; Output; Paper; Pathogen detection; Patients; Pattern; Physicians; Play; Population; Powassan virus; Publishing; Reading; Reagent; Refrigeration; Reporting; Reproduction; Resources; Rickettsia; Rickettsia Infections; Rickettsia rickettsii; Risk; Rocky Mountain Spotted Fever; Role; Sampling; Serotyping; Services; Ships; Signal Transduction; Sterilization; Testing; Tick-Borne Diseases; Ticks; Time; Tube; Uncertainty; United States National Institutes of Health; United States Public Health Service; Urine; Vector-transmitted infectious disease; Virus; Work; Zika Virus; ammonium hydroxide; base; biosafety level 2 facility; blind; chikungunya; college; cost; design; genetic information; human disease; human pathogen; mosquito-borne; mosquito-borne pathogen; pathogen; pathogen genome; response; success; tick-borne; tick-borne pathogen; vector

Point-of-sampling detection of Lyme and other tick-borne diseases Foundation for Applied Molecular Evolution Steven A. Benner ABSTRACT Lyme disease is today the most prevalent vector-borne disease in the US, and ticks in general are the most significant vectors for human pathogens worldwide, including Borrelia, Babesia, and Rickettsia species. For Lyme disease, the CDC confirmed only 28,000 cases in a population that might actually hold 300,000. This severe underdiagnosis is due to: (a) expensive and spotty environmental surveillance, yielding insufficient patient and physician awareness, and (b) inadequate molecular tests; those exploiting antibodies can give 86% false negatives. Further, to order a molecular test (at $260), a physician must "guess" that a patient suffers from one of these diseases. As single pathogen-single tests are the norm in the FDA-regulated diagnostics space, these guesses must expand (each $200) to other candidate pathogens, one at a time. Not surprisingly, aggressive advocacy groups have arisen to counter (what they feel to be) inadequate CDC/NIH response to this disease threat. This R21 project will deliver a molecular test that, in one step in the hands of uncertified users, identifies a complete panel of all tick-borne pathogens, for less than $1.00, and within 30 minutes. The assay will apply to ticks directly, either trapped or pulled off a pet or a child. It will be useable by college nurses, doctors in their offices, public health service staff, and even laypeople, to examine ticks. After this R21 project is complete, the data will set the stage for a human diagnostic kit that can be FDA-approved to apply to patient samples. Although the Benner lab has never before worked with tick-borne diseases, this proposal has a high chance for success, as it has been de-risked by work, just published, where the lab delivered a kit that detects dengue, chikungunya, and/or Zika in a single assay, in 30 minutes, in a single mosquito carcass. The sample is rendered free of biohazard by treatment with ammonia (similar to Windex®) and ethanol, and dropped into a pre- prepared tube containing dry reagents, shipped without refrigeration. Alternatively, urine or blood may be used. This kit is being used today to monitor the presence of pathogens in trapped mosquitoes, and shipped to China and India, where it detects dengue and chikungunya in human urine and blood. This illustrates our strategy of developing unregulated tests to support introduction of tests for patient use. The deliverable will be benchmarked on real ticks containing real pathogens, provided by the laboratory of Maria Diuk-Wasser at Columbia. This will include blind testing on ca. 50 samples. The premises are: (a) The NH3-EtOH disruption is sufficient to release detectable amounts of pathogen target; (b) multiplexing in a reverse-LAMP architecture can be increased from 3 to ~10 to cover all tick targets; and (c) AEGIS in displace- able probes will support a "patterned readout" for different pathogens, and a control. The authentication of the reagents will be assured by making the deliverable available to the public, as is being done for mosquito- borne pathogen detection.

莱姆病和其他蜱传疾病的采样点检测 应用分子进化基金会 史蒂文·A·本纳 抽象的 莱姆病是当今美国最流行的媒介传播疾病，蜱虫是最常见的媒介传播疾病 全世界人类病原体的重要载体，包括疏螺旋体、巴贝虫和立克次体。 莱姆病期间，美国疾病控制与预防中心 (CDC) 在实际可能有 30 万人口的人群中仅确认了 28,000 例病例。 严重漏诊的原因是： (a) 昂贵且不稳定的环境监测，导致患者和医生意识不足，以及 (b) 分子检测不充分；利用抗体的检测可能会出现 86% 的假阴性。 此外，要进行分子检测（售价 260 美元），医生必须“猜测”患者患有其中一种疾病 由于单一病原体单一测试是 FDA 监管的诊断领域的常态，因此这些猜测。 必须扩展到其他候选病原体（每个 200 美元），一次一种，积极宣传，这并不奇怪。 一些团体已经起来反对（他们认为的）CDC/NIH 对这种疾病威胁的反应不足。 该 R21 项目将提供一种分子测试，在未经认证的用户手中一步即可识别出 所有蜱传病原体的完整检测，价格不到 1.00 美元，并且可在 30 分钟内进行检测。 直接蜱虫，无论是被困住还是从宠物或孩子身上拉下来，护士学院、医生都可以使用。 R21 项目完成后，办公室、公共卫生服务人员甚至外行人员都可以检查蜱虫。 数据将为人类诊断试剂盒奠定基础，该试剂盒可以经 FDA 批准应用于患者样本。 尽管本纳实验室以前从未研究过蜱传疾病，但这一提议有很高的机会 为了成功，因为它已经通过刚刚发表的工作降低了风险，实验室提供了一个检测登革热的试剂盒， 基孔肯雅热和/或寨卡病毒可在 30 分钟内在单个蚊子尸体中进行一次检测。 通过氨水（类似于 Windex®）和乙醇处理，无生物危害，并放入预干燥器中。 装有干燥试剂的准备好的试管，无需冷藏即可运输。或者，也可以使用尿液或血液。 该套件目前用于监测被困蚊子中是否存在病原体，并运往。 中国和印度在人类尿液和血液中检测到登革热和基孔肯雅热，这说明了我们的情况。 开发不受监管的测试以支持引入供患者使用的测试的策略。 该交付成果将以包含真实病原体的真实蜱虫为基准，由实验室提供 哥伦比亚的 Maria Diuk-Wasser 将包括对大约 50 个样本进行盲测。 NH3-EtOH 破坏足以释放可检测量的病原体靶标；(b) 在 反向 LAMP 架构可以从 3 个增加到约 10 个，以覆盖所有蜱目标；以及 (c) 位移中的 AEGIS； 能够的探针将支持不同病原体的“图案读出”和控制的验证。 试剂将通过向公众提供交付品来保证，就像针对蚊子所做的那样 携带病原体检测。