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

基本信息

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
项目类别：
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

项目摘要

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，但报道不足。据估计，每年有数千万人受到感染，如果不在感染早期进行治疗，可能会导致严重的并发症，甚至可能致命。钩端螺旋体病是通过皮肤或粘膜接触受感染动物的尿液而感染的。最常见的是，这种情况是由于接触受污染的水体或土壤而发生的。世界卫生组织列出了有感染风险的人群，包括养牛和养猪的农民、兽医、屠夫和屠宰场工人、在雨坑或泥巴中玩耍的儿童，以及从事游泳、划船、游泳等娱乐活动的人。洞穴探险或钓鱼。简而言之，很少有人没有感染钩端螺旋体病的风险。然而，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见。因此，迫切需要一种快速、准确的现场诊断工具，用于检测可能接触尿液污染的土壤和水体的患者的致病性钩端螺旋体。为了应对这一挑战，Luna Innovations Incorporated 和 Rapid Pathogen Screening, Inc. 建议开发基于免疫亲和力的钩端螺旋体检测方法，使用严格调控的蛋白水解作用来提供酶信号放大。拟议的第一阶段计划将提供一种高度灵敏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来自放大级联，最终导致作用于信号分子的酶的完全激活。由于信号是由放大的酶活性产生的，因此可以想象，临床样品中的单个致病颗粒将产生相同水平的蛋白水解活性，因此与含有数千种病原体的样品产生的信号相同的强信号。兴趣。因此，Luna 的病原体诱导信号放大平台应该能够产生非常灵敏且快速的现场诊断分析。对于这一第一阶段计划，Luna 将生产、纯化和表征新型检测试剂，并展示诊断致病性钩端螺旋体检测的灵敏度和选择性。第二阶段计划将重点是将基于溶液的测定转换为侧向流动格式，以实现用户友好的即时测定，并验证该测定的临床应用。公共卫生相关性：钩端螺旋体病是一种因皮肤或粘膜接触受感染动物的尿液而引起的感染，被认为是世界范围内最常见的人畜共患病。据估计，每年有数千万人受到影响，死亡率在 5% 至 25% 之间。钩端螺旋体病可以用抗生素治疗，但早期诊断对于抗生素治疗的有效性是必要的。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见。 Luna Innovations Incorporated 和 Rapid Pathogen Screening, Inc. (RPS) 提议开发一种灵敏且选择性的即时诊断检测方法，用于直接检测临床样本中的致病性钩端螺旋体，以便在全球资源有限的地区部署。