Point-of-Care Diagnostic Assay for Direct Detection of Pathogenic Leptospira

直接检测致病性钩端螺旋体的即时诊断分析

• 批准号: 7998343
• 负责人: Michael J Danilich
• 金额: $ 11.8万
• 依托单位: LUNA INNOVATIONS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-02 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7998343
• 关键词: Abattoirs; Address; Affect; Agglutination Tests; Animals; Antibiotic Therapy; Antibiotics; Antibodies; Bacillus (bacterium); Bacteria; Biological Assay; Biological Warfare; Caspase; Cattle; Child; Chimera organism; Clinical; Contracts; Detection; Development; Diagnosis; Diagnostic; Disease Outbreaks; Early Diagnosis; Enzyme Activation; Familiarity; Family suidae; Fishes; Francisella tularensis; Funding; Government; In Vitro; Incubated; Infection; Influenza; Laboratories; Lateral; Lead; Left; Leptospira; Leptospirosis; Libraries; Methods; Microbiology; Microscopic; Mucous Membrane; Mycoses; Pathogen detection; Patients; Peptide Hydrolases; Peptides; Persons; Phage Display; Phase; Physicians; Play; Proteins; Proteolysis; Rain; Reagent; Resources; Risk; Sampling; Screening procedure; Sensitivity and Specificity; Serum; Signal Transduction; Signaling Molecule; Skin; Soil; Solutions; Staging; Surface Antigens; Swimming; System; Time; Urine; Veterinarians; Viral; Virus Diseases; Water; Work; World Health Organization; Zoonoses; assay development; base; boating; caspase-3; clinical application; design; expression vector; farmer; innovation; interest; mortality; novel; particle; pathogen; phase 1 study; point of care; point-of-care diagnostics; programs; prototype; public health relevance; tool; user-friendly

DESCRIPTION (provided by applicant): Leptospirosis is the most common zoonosis yet it is underreported. It is estimated to affect tens of millions annually and if not treated early in the infection can lead to severe complications and can be fatal. Leptospirosis is contracted through skin or mucous membrane contact with urine of infected animals. Most often this occurs by contact with contaminated bodies of water or soil. The World Health Organization lists the groups of people at risk of the infection ranging from cattle and pig farmers to veterinarians, butchers and slaughterhouse workers to children playing in rain puddles or mud, as well as people engaging in recreational activities such as swimming, boating, caving or fishing. In short, there are few people who are not at risk of leptospirosis infection. However, because the initial presentation of the infection is often that of non-specific viral infections, such as influenza, early diagnosis and treatment is infrequent. Thus, there is a critical need for a rapid and accurate point-of-care diagnostic tool for detecting pathogenic Leptospira in patients potentially exposed to urine contaminated soils and bodies of water. To address this challenge, Luna Innovations Incorporated and Rapid Pathogen Screening, Inc. propose to develop immunoaffinity-based Leptospira detection assays using tightly-regulated proteolysis to provide enzymatic signal amplification. The proposed Phase I program will deliver a highly sensitive and easy-to-use rapid diagnostic tool for direct detection of pathogenic Leptospira. A key feature of this platform design is that the signal results from an amplification cascade culminating in complete activation of the enzymes that act upon the signaling molecule. Because the signal is generated by the amplified enzymatic activity, it is conceivable that a single pathogenic particle in the clinical sample would generate the same level of proteolytic activity, and therefore the same robust signal as that generated by a sample containing thousands of the pathogens of interest. Thus, Luna's pathogen-induced signal amplification platform should produce very sensitive as well as rapid point-of-care diagnostic assays. For this Phase I program, Luna will produce and purify and characterize novel assay reagents and demonstrate sensitivity and selectivity of the assay for diagnosis of pathogenic Leptospira. The Phase II program will focus on converting the solution-based assay into the lateral flow format for a user friendly point-of-care assay as well as validating the assay for clinical applications. PUBLIC HEALTH RELEVANCE: Leptospirosis, an infection resulting from skin or mucous membrane contact with urine of infected animals, is considered the most common zoonosis worldwide. It is estimated to affect tens of millions annually with mortality rates ranging from 5 to 25%. Leptospirosis is treatable with antibiotics, however early diagnosis is necessary for antibiotic therapy to be effective. Because the initial presentation of the infection is often that of non-specific viral infections, such as influenza, early diagnosis and treatment is infrequent. Luna Innovations Incorporated and Rapid Pathogen Screening, Inc. (RPS) propose to develop a sensitive and selective point-of- care diagnostic assay for the direct detection of pathogenic Leptospira in clinical samples for deployment in resource-limited regions of the globe.

描述（由申请人提供）：钩端螺旋体病是最常见的人畜共患病。据估计，它每年会影响数千万，如果在感染的早期未接受治疗，可能会导致严重的并发症，可能是致命的。钩端螺旋体通过皮肤或粘膜接触与感染动物的尿液接触。大多数情况下，这是通过与受污染的水或土壤接触而发生的。世界卫生组织列出了有感染风险的人群，从牛，养猪者到兽医，屠夫和屠宰场工人到在雨水水坑或泥浆中玩耍的儿童，以及从事游泳，划船，划船，划船，划船，划船，洞穴或钓鱼。简而言之，很少有人没有患钩端螺旋体病的风险。但是，由于感染的最初表现通常是非特异性病毒感染（例如流感），因此早期诊断和治疗很少。因此，在潜在暴露于尿液受污染的土壤和水体的患者中，需要快速准确的护理诊断工具来检测病原性钩端螺旋体。为了应对这一挑战，Luna Innovations Incorporated and Rapid病原体筛查Inc.建议使用紧密调节的蛋白水解开发基于免疫亲和力的Leptospira检测试验，以提供酶促信号扩增。拟议的I期计划将提供高度敏感且易于使用的快速诊断工具，用于直接检测致病性钩端螺旋体。该平台设计的一个关键特征是，信号是由放大级联反对在信号分子上的酶的完全激活中引起的。因为信号是由扩增的酶促活性产生的，所以可以想象的是，临床样品中的单个致病粒子将产生相同水平兴趣。因此，Luna病原体诱导的信号扩增平台应产生非常敏感和快速的护理诊断测定法。对于此I阶段计划，Luna将产生，纯化并表征新颖的测定试剂，并证明对诊断致病性瘦心的测定的敏感性和选择性。第二阶段计划将重点介绍将基于溶液的测定法转换为侧向流动格式，以进行用户友好的护理测定法，并验证用于临床应用的测定法。 公共卫生相关性：钩端螺旋体病，是皮肤或粘膜与感染动物尿液接触引起的感染，被认为是全球最常见的人畜共患病。据估计，每年数千万的死亡率范围为5％至25％。钩端螺旋体病可以用抗生素治疗，但是抗生素治疗的早期诊断是有效的。由于感染的最初表现通常是非特异性病毒感染（例如流感），因此早期诊断和治疗很少。 Luna Innovations Incormated and Rapid病原体筛查Inc.（RPS）建议开发一种敏感和选择性的诊断测定，以直接检测临床样本中的致病性Leptospira，以在全球资源有限的地区部署中部署。