Hybrid ELISA: Simple and specific one-tier assay for Lyme disease

混合 ELISA：针对莱姆病的简单而特异的一层检测

• 批准号: 10758919
• 负责人: Andrew E. Levin
• 金额: $ 102.5万
• 依托单位: KEPHERA DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-06 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10758919
• 关键词: Accounting; Algorithms; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Antigens; Area; Attention; Bacterial Antigens; Bacterial Infections; Binding; Biological Assay; Blood; Borrelia; Borrelia burgdorferi; Chronic Disease; Clinical; Clinical Research; Communicable Diseases; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Enzyme-Linked Immunosorbent Assay; Epitopes; European Union; Evaluation; Exanthema; Eye; Formulation; Geography; Government; Grant; Growth; Hybrids; Immunochemistry; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Laboratories; Legitimacy; Location; Lyme Disease; Lyme disease diagnosis; Marketing; Medical; Methodology; Methods; Midwestern United States; Nature; New England; Order Spirochaetales; Patients; Peptides; Performance; Persons; Phase; Physicians; Predictive Value; Preparation; Procedures; Process; Proteins; Protocols documentation; Public Health; Reagent; Recombinant Proteins; Recommendation; Recording of previous events; Reproducibility; Research Design; Rest; Risk; Risk Factors; Running; Sampling; Screening Result; Sensitivity and Specificity; Serology; Serology test; Serum; Services; Site; Specificity; Step Tests; Symptoms; Test Result; Testing; Tick-Borne Diseases; Time; Translations; United States; Validation; Vector-transmitted infectious disease; Western Blotting; antibody detection; assay development; clinical diagnosis; commercial launch; cost; cross reactivity; diagnostic assay; erythema migrans; in-vitro diagnostics; manufacture; novel; prospective; prototype; research clinical testing; screening; synthetic peptide

Abstract Lyme disease, caused by infection with the spirochete Borrelia burgdorferi or closely related species, is the most common vector-borne disease in the United States, accounting for nearly 500,000 infections per year according to recent public health estimates. It is also one of the few infectious diseases that requires a two-step laboratory testing protocol, comprising a screening assay, typically ELISA, followed by a confirmatory immunoblot. This protocol is followed where the disease has progressed beyond the hallmark erythema migrans rash, or the rash cannot be identified, and symptoms or history suggest Lyme disease. The two-step protocol is a result of the poor specificity of most Lyme screening tests, which lead to a high false positive rate. This in turn is due to the cross-reactive nature of many bacterial antigens, combined with the limitations of conventional ELISA immunochemistry. In principle, a confirmatory result must be obtained prior to initiating treatment, but in practice, the multi-day delay that is often incurred in this process frequently leads physicians to prescribe antibiotic treatment in the absence of definitive lab results. As the vast majority of the more than three million annual Lyme tests in the U.S. are carried out on individuals who are ultimately found not to have Lyme disease, this practice leads to the unnecessary prescription and use of antibiotics, contributing to the growth of antibiotic resistance which has become a significant threat to public health. A first step test with significantly higher specificity would avoid the need for a second step test and enable clinicians to make informed treatment decisions in a timely manner, based on credible test results. This project is aimed at bringing about a significant change in medical practice by reducing Lyme testing from the current two-step process to a one-step process. To achieve this objective, we have developed a novel ELISA methodology for Lyme antibody detection that enables exceptionally high assay specificity. The test is based on well-established, highly specific and sensitive Borrelia antigens in a unique immunochemical format. The novel ELISA immunochemistry eliminates almost all non-specific reactivity, yielding results comparable in specificity but higher in sensitivity than those obtained with the conventional two-tier testing protocol. Consequently, this assay promises to deliver a one-step testing solution for Lyme disease, at a time when alternatives to the original two-step method are gaining legitimacy at the scientific as well as regulatory level. In Phase I, we developed a prototype ELISA assay, proving feasibility by demonstrating higher sensitivity and equivalent specificity on retrospective samples comprising Lyme patients and controls, in comparison with two- tier testing results. In Phase II, we will complete development of the assay into a commercial product, carry out prospective and retrospective clinical studies and submit a 510(k) application to FDA for use of the ELISA as a one-step test for Lyme disease, enabling commercial launch upon approval.

摘要 莱姆病是由伯氏疏螺旋体或近缘物种感染引起的 据报道，美国常见的媒介传播疾病每年造成近50万人感染 与最近的公共卫生评估结果相吻合。它也是为数不多的需要两步实验室的传染病之一。 检测方案，包括筛查试验，通常是酶联免疫吸附试验，然后是确认性免疫印迹。这 如果疾病的进展超出了标志性红斑、游走性皮疹或皮疹，则按照协议进行 无法辨认，症状或病史表明是莱姆病。两个步骤的协议是 大多数莱姆筛查试验的特异性较差，导致假阳性率较高。这反过来又是由于 许多细菌抗原的交叉反应性质，结合传统酶联免疫吸附试验的局限性 免疫化学。原则上，在开始治疗之前必须获得确诊结果，但在实践中， 在这个过程中经常会出现数天的延误，这经常导致医生开抗生素处方。 在没有确切实验室结果的情况下进行治疗。作为每年300多万莱姆中的绝大多数 在美国，测试是在最终被发现没有莱姆病的个人身上进行的，这种做法 导致不必要的抗生素处方和使用，导致抗生素耐药性的增长 这已经成为对公共健康的重大威胁。具有明显更高特异性的第一步测试将 避免第二步测试的需要，使临床医生能够及时做出知情的治疗决定 方式，基于可信的测试结果。 该项目旨在通过减少莱姆测试来显著改变医疗实践 从目前的两步过程转变为一步过程。为了实现这一目标，我们开发了一种新型的酶联免疫吸附试验 莱姆抗体检测方法，可实现极高的检测特异性。这项测试是基于 以独特的免疫化学形式对成熟、高度特异和敏感的疏螺旋体抗原进行研究。这个 新型的ELISA免疫化学消除了几乎所有的非特异性反应，产生了与 特异度高，但灵敏度高于传统的两层检测方法。 因此，这项检测有望为莱姆病提供一步检测解决方案，而此时 最初的两步法的替代方法在科学和监管层面都获得了合法性。 在第一阶段，我们开发了一个原型ELISA法，通过展示更高的灵敏度和 对包括莱姆病患者和对照的回溯性样本的等同特异度，与两个- 分层测试结果。在第二阶段，我们将完成该检测方法的商业化开发，进行 前瞻性和回顾性的临床研究，并向FDA提交510(K)申请，以将该酶联免疫吸附试验用作 莱姆病一步检测，批准后即可投入商业使用。