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.

抽象的 莱姆病是由伯氏疏螺旋体或密切相关的物种感染螺旋体引起的，是最常见的疾病。 据统计，美国常见的媒介传播疾病每年造成近 50 万例感染 最近的公共卫生估计。它也是少数需要两步实验室的传染病之一 测试方案，包括筛选测定（通常是 ELISA），然后是验证性免疫印迹。这 当疾病进展超出标志性游走性红斑皮疹或皮疹时，应遵循治疗方案 无法识别，症状或病史提示莱姆病。两步协议的结果是 大多数莱姆病筛查测试的特异性较差，导致假阳性率较高。这又是由于 许多细菌抗原的交叉反应性质，加上传统 ELISA 的局限性 免疫化学。原则上，在开始治疗之前必须获得确认结果，但实际上， 在此过程中经常出现多天的延迟，经常导致医生开出抗生素 在没有明确的实验室结果的情况下进行治疗。每年超过 300 万莱姆病患者中的绝大多数 在美国，测试是针对最终发现没有患有莱姆病的个体进行的，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 这已成为对公众健康的重大威胁。具有显着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明智的治疗决定 方式，基于可信的测试结果。 该项目旨在通过减少莱姆病检测来给医疗实践带来重大改变。 将当前的两步过程改为一步过程。为了实现这一目标，我们开发了一种新型 ELISA 莱姆病抗体检测方法可实现极高的检测特异性。测试基于 以独特的免疫化学形式对成熟、高度特异性和敏感的疏螺旋体抗原进行检测。这 新型 ELISA 免疫化学消除了几乎所有非特异性反应，产生的结果与 与传统的两层测试方案相比，特异性高，但灵敏度更高。 因此，该检测有望为莱姆病提供一步式检测解决方案 原始两步法的替代方案正在科学和监管层面获得合法性。 在第一阶段，我们开发了一种原型 ELISA 检测方法，通过展示更高的灵敏度和 与两种方法相比，包括莱姆病患者和对照的回顾性样本具有同等的特异性 等级测试结果。在第二阶段，我们将完成将该检测方法开发为商业产品，进行 前瞻性和回顾性临床研究，并向 FDA 提交 510(k) 申请，以使用 ELISA 作为检测方法 莱姆病的一步测试，经批准即可商业化。