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: 9886194
• 负责人: Andrew E. Levin
• 金额: $ 29.98万
• 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 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9886194
• 关键词: Accounting; Adverse effects; Amoxicillin; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Antigens; Bacterial Antigens; Bacterial Infections; Binding; Biological Assay; Blood; Blood donor; Borrelia; Borrelia burgdorferi; Cells; Centers for Disease Control and Prevention (U.S.); Chronic Disease; Clinical; Clinical Research; Clinical Sensitivity; Communicable Diseases; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Doxycycline; Enzyme-Linked Immunosorbent Assay; Epitopes; Exanthema; Eye; Formulation; Geographic Locations; Gold Colloid; Grant; Growth; Horseradish Peroxidase; Hybrids; Immunochemistry; Immunoglobulin G; Immunoglobulin M; In Vitro; Individual; Infection; Laboratories; Legitimacy; Lyme Disease; Medical; Methodology; Methods; Nature; Order Spirochaetales; Outcome; Patients; Peptides; Performance; Peroxidases; Phase; Physicians; Plasma; Polymers; Process; Proteins; Protocols documentation; Public Health; Reagent; Recombinant Proteins; Recording of previous events; Rest; Risk; Sampling; Screening Result; Sensitivity and Specificity; Serologic tests; Serological; Serum; Short-Term Course; Signal Transduction; Specificity; Statistical Data Interpretation; Step Tests; Symptoms; Test Result; Testing; Time; United States; Vector-transmitted infectious disease; base; cross reactivity; erythema migrans; novel; particle; prospective; prototype; 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 over 300,000 infections per year according to 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 results in 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 for this purpose leads to physicians frequently prescribing antibiotic treatment in the absence of definitive lab results. As the vast majority of the roughly three million annual Lyme tests in the U.S. are carried out on individuals who do not 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 both sensitivity and specificity to 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 this Phase I project, we will develop a prototype ELISA assay, proving feasibility by demonstrating sensitivity and specificity on retrospective samples comprising Lyme patients and controls, in comparison with two-tier testing results. In Phase II, we will complete development of a commercial product, carry out prospective and retrospective clinical studies and submit an application to FDA for in vitro diagnostic claims, allowing commercial launch upon approval.

摘要 莱姆病是由螺旋体伯氏疏螺旋体或密切相关的物种感染引起的， 这是美国最常见的病媒传播疾病，每年有30多万人感染 根据公共卫生评估。它也是为数不多的需要两步走的传染病之一 实验室测试方案，包括筛选测定，通常为ELISA，随后是确认性测定， 免疫印迹。如果疾病进展超过标志性红斑，则遵循该方案 游走性皮疹，或皮疹无法识别，症状或病史提示莱姆病。两步 方案是由于大多数莱姆筛查试验的特异性差，导致高假阳性 率这反过来又是由于许多细菌抗原的交叉反应性质，加上限制 常规ELISA免疫化学。原则上，必须在启动前获得确证性结果 治疗，但在实践中，多天的延误，往往是为此目的导致医生 在没有明确的实验室结果的情况下经常开抗生素治疗。作为绝大多数的 在美国，每年大约有300万例莱姆病检测是在没有莱姆病的人身上进行的 疾病，这种做法导致不必要的处方和使用抗生素，有助于增长， 抗生素耐药性已成为对公众健康的重大威胁。第一步测试， 更高的特异性将避免第二步测试的需要，并使临床医生能够做出明智的治疗 根据可靠的测试结果及时做出决定。 该项目旨在通过减少莱姆病检测， 将目前的两步法改为一步法。为了实现这一目标，我们开发了一种新的 用于莱姆病抗体检测的ELISA方法能够实现极高的检测特异性。测试 基于以独特的免疫化学形式建立的高度特异性和敏感性的疏螺旋体抗原。 新的ELISA免疫化学消除了几乎所有的非特异性反应，产生的结果与 灵敏度和特异性均优于用常规的两级测试方案获得的那些。因此，委员会认为， 这种检测方法有望为莱姆病提供一步式检测解决方案， 最初的两步法在科学和监管层面上获得了合法性。 在这个第一阶段的项目中，我们将开发一个原型ELISA检测，通过证明灵敏度来证明可行性 和特异性的回顾性样本包括莱姆病患者和对照组，与两级相比， 测试结果在第二阶段，我们将完成商业产品的开发，开展前瞻性和 回顾性临床研究，并向FDA提交体外诊断声明申请， 经批准后进行商业发射。