Development of Molecular Diagnostic Test for Early Onset of Lyme disease

莱姆病早期发作的分子诊断测试的开发

• 批准号: 9473695
• 负责人: STEVEN E SCHUTZER
• 金额: $ 98.84万
• 依托单位: BIOSCIENCE DEVELOPMENT, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-07 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9473695
• 关键词: Antibiotic Therapy; Antibiotics; Antibodies; Antibody Response; Area; Bacteria; Benchmarking; Biological; Biological Assay; Blood; Blood Tests; Blood Volume; Blood specimen; Borrelia; Borrelia burgdorferi; Centers for Disease Control and Prevention (U.S.); Clinical; Communicable Diseases; Consult; DNA; DNA copy number; Data; Deposition; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Early Diagnosis; Economic Burden; Enzyme-Linked Immunosorbent Assay; Exanthema; Eye; Feeling; Goals; Health system; Heart; Hour; Infection; Joints; Laboratories; Left; Lyme Disease; Methods; Microbe; Molecular Diagnostic Testing; Nervous system structure; Patients; Phase; Public Health; Publishing; Research Design; Sampling; Sensitivity and Specificity; Serologic tests; Sore Throat; Specificity; Spottings; Symptoms; Technology; Testing; Ticks; Time; Tissues; United States; Urinary tract infection; West Nile virus; Western Blotting; Whole Blood; Work; base; burden of illness; case control; clinically relevant; common symptom; cost; diagnostic assay; disabling symptom; disorder control; early onset; erythema migrans; flu; impression; novel strategies; nucleic acid detection; pathogen; prevent; seroconversion; skin lesion

Project Summary/Abstract There is no validated assay to reliably detect the early onset of the Lyme disease infection at the time when antibiotic therapy has the best chance of a cure without sequelae. Diagnosis is often difficult because most symptoms are often nebulous, with the telltale “bulls-eye” rash only appearing and clearly recognizable in approximately half of cases. The standard types of infectious diagnostic tests – both culture and serology – require >3 weeks for a positive result after the onset of infection with Lyme disease and serology only indicates exposure but not active infection. This introduces a significant delay in treatment. While PCR for B. burgdorferi DNA in the blood has potential for a rapid, accurate result, past tests have not had sufficient sensitivity because of too few bacteria and low copy numbers of DNA in the blood. We have developed an approach that “enlarges the targetability”, by 160,000 fold, thereby increasing the sensitivity of PCR test for B. burgdorferi. Our data show we are successfully accomplishing this by: using a larger amount of blood to start (10 fold); then selectively multiplying the target by using Borrelia-directed primers prior to PCR (2000 fold); and then probing the product for the B. burgdorferi on a multi-loci PCR platform (8 fold). To assure reliability and acceptance of a new test based on this approach, we formally consulted with the FDA from the beginning. We obtained a Qsub (pre-IDE) approval with guidance as to what we need for a cleared test. During this proposal we will continue to test blood samples (from eventually confirmed Lyme cases and control subjects) with our PCR-based assay at the time the patient presents to the doctor in order to prove the assay can detect the B. burgdorferi infection, earlier and with greater sensitivity, than the FDA benchmark of two-tiered serology taken at the same time. If successful, at the number agreed on with the FDA, this Phase II should result in a test that has FDA clearance. Our goal is deliver a functional and affordable diagnostic assay for Early Onset Lyme Disease that has superior sensitivity and specificity compared to two-tiered serology. The test will permit immediate diagnosis well before seroconversion or cultures become positive so treatment can be started right away.

项目摘要/摘要 没有经过验证的测定可靠地检测到当时莱姆病的早期发作 当抗生素疗法具有无后遗症治愈的最佳机会时。诊断通常很困难 因为大多数符号通常都是模糊的，所以出现的“牛眼”皮疹只会出现，并且 在大约一半的情况下，显然可以识别。 传染性诊断测试的标准类型（无论是培养还是血清学）需要> 3周 莱姆病和血清学感染发作后的积极结果仅表明暴露，但 没有主动感染。这引入了严重的治疗延迟。而B. burgdorferi的PCR 血液中的DNA具有快速，准确的结果的潜力，过去的测试还没有足够 敏感性是因为细菌太少和血液中DNA的低拷贝数。我们有 开发了一种“扩大目标性”的方法，提高了160,000倍，从而增加了 PCR测试对B. burgdorferi的敏感性。我们的数据显示，我们正在成功完成此操作： 使用更多的血液开始（10倍）；然后通过使用 PCR之前的伯氏指导引物（2000倍）；然后探测B. burgdorferi的产品 在多层PCR平台上（8倍）。 为了确保基于这种方法的新测试的可靠性和接受，我们正式咨询了 FDA从一开始。我们获得了有关我们什么的QSUB（IDE）批准 需要清除测试。在此提案中，我们将继续测试血液样本（最终 确认的莱姆病病例和对照受试者）在患者时使用我们的基于PCR的测定法 为了证明该测定法可以检测到B. burgdorferi感染，并且 比同时获得两层血清学的FDA基准的灵敏度更高。如果 成功，按照与FDA达成的数字，该阶段II应该导致具有FDA的测试 清除。 我们的目标是为早期发作莱姆病提供功能和负担得起的诊断测定法 与两层血清学相比，具有较高的灵敏度和特异性。测试将允许 在血清转化或培养物变为阳性之前，可以立即进行诊断，因此可以治疗 立即开始。