Highly-sensitive, rapid and low cost plasmonic assay platform for Lyme disease diagnosis

用于莱姆病诊断的高灵敏度、快速且低成本的等离子体检测平台

• 批准号: 10546574
• 负责人: Nathaniel Charles Cady
• 金额: $ 25.96万
• 依托单位: CIENCIA, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-12 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546574
• 关键词: 3D Print; Acute; Address; Adoption; Advisory Committees; Algorithms; Antibiotic Therapy; Antibodies; Antibody Response; Antibody titer measurement; Antigens; Benchmarking; Biocompatible Materials; Biological Assay; Blinded; Blood; Blood Volume; Borrelia; Borrelia burgdorferi; Businesses; Centers for Disease Control and Prevention (U.S.); Chronically Ill; Clinical; Complex; Comprehensive Health Care; Computer software; Consultations; Consumption; Coupled; Data Analyses; Detection; Development; Device Designs; Devices; Diagnostic; Disease; Early Diagnosis; Endemic Diseases; Environment; Enzyme-Linked Immunosorbent Assay; European; Fluorescence; Funding; Goals; Gold; Health; Immune response; Immunity; Immunoglobulin Class Switching; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Institutes; Investments; Laboratories; Liquid substance; Lyme Disease; Lyme disease diagnosis; Lyme disease diagnostic; Methods; Monitor; Multiple Sclerosis; Needs Assessment; Order Spirochaetales; Patients; Phase; Preparation; Price; Process; Protocols documentation; Publishing; Reagent; Research; Research Personnel; Rheumatoid Arthritis; SARS-CoV-2 antibody; SARS-CoV-2 infection; Sampling; Sensitivity and Specificity; Serology; Serology test; Serum; Signal Transduction; Silicon; Site; Small Business Technology Transfer Research; Specificity; Spottings; Streptavidin; Syphilis; System; Technology; Test Result; Testing; Tick-Borne Diseases; Time; United States; Vaccination; Variant; Vector-transmitted infectious disease; Western Blotting; Work; base; clinically relevant; cohort; consumer demand; cost; cross reactivity; design; detection limit; diagnostic algorithm; diagnostic assay; diagnostic strategy; diagnostic value; flexibility; health care service; improved; interest; next generation; plasmonics; point of care; programs; prototype; response; screening; sensor; success

Project Summary/Abstract Lyme disease (LD) is the most common vector-borne illness in the United States and represents a considerable diagnostic challenge. 1-3 The current benchmark for LD diagnosis is the standard two-tiered test (STTT), although the modified two-tier test (MTTT) is rapidly becoming an acceptable alternative. 6 These assays are highly specific but have poor sensitivity, especially early in the infection. 6, 7 Traditional testing is limited by each patient's heterogenous immune response to Borrelia burgdorferi (Bb) antigens. While the paired ELISAs of the MTTT are sensitive and simple to perform, some patients respond weakly to screening reagents. The STTT addresses variation in humoral response by combining an initial ELISA with a confirmatory IgG/IgM Western Blot (WB), yet WB sensitivity is modest against any single antigen. This is especially true with IgM results, which also lack the specificity enhancement associated with class switching. 8 Given these limitations, the ideal serologic assay will combine features of ELISA and WB, but will provide quantitative and specific results against all major LD antigens. Using current technologies, this approach is cost prohibitive. Ciencia Inc. has developed a low-cost, highly-sensitive, fluorescence-based assay platform that supports exactly this type of quantitative multi-antigen serologic Lyme assay. 4, 9, 10 Dr. Cady at SUNY Poly has used our grating-coupled fluorescent plasmonics (GC-FP) assay platform to define antibody responses against a suite of Bb antigens. This preliminary work demonstrates superior sensitivity/specificity compared to STTT (90%/100% vs 60%/100%; n=34 validated samples) and an earlier time to result. Our approach offers a linear response across the full dynamic range, a rapid time to result, low reagent costs. Antigen spots may be added for a negligible cost, so a broader tickborne disease (TBD) panels could be performed for the price of one ELISA. With this Phase I STTR, Ciencia will partner with Dr. Cady (SUNY Polytechnic Institute) and Dr. Strle (Wadsworth Center) to build upon our preliminary self-funded exploration of Lyme serodiagnostics. We aim to validate our preliminary results using a larger set of well-characterized LD samples and appropriate controls from the CDC and NY State DoH, including an estimate of time to positivity using samples collected longitudinally. In parallel, Ciencia aims to improve sample handling and overall ease of use by implementing a cartridge-based system and automated processing that will make GC-FP a viable alternative to methods such as the STTT and MTTT, both of which are cumbersome and time consuming. The ultimate goal is to provide a rapid, easy-to-use, and highly accurate test that can be used outside traditional clinical laboratories. Based on the success of Phase I, the Phase II effort would optimize the test for diagnosis of Lyme disease caused by European Borrelia strains and expand it to other tickborne diseases. Phase II would also permit design refinements and initiation of the FDA approval process.

项目总结/摘要 莱姆病（LD）是美国最常见的病媒传播疾病， 诊断上的巨大挑战。1-3目前LD诊断的基准是标准的两层测试 （STTT），虽然修改后的两级测试（MTTT）正在迅速成为一个可接受的替代方案。6这些 检测是高度特异性的，但灵敏度差，尤其是在感染早期。六、七 传统的测试是有限的，每个病人的异质性免疫反应，伯氏疏螺旋体（Bb） 抗原虽然MTTT的配对ELISA敏感且操作简单，但一些患者对MTTT有反应。 对筛选试剂的敏感性较弱。STTT通过结合初始的 ELISA与确证性IgG/IgM蛋白质印迹法（WB），但WB对任何单一抗原的灵敏度都是中等的。 IgM结果尤其如此，IgM结果也缺乏与分类相关的特异性增强。 切换考虑到这些局限性，理想的血清学检测将结合ELISA和WB的联合收割机特征，但 提供针对所有主要LD抗原的定量和特异性结果。利用现有技术， 这种方法成本过高。 Cipher Inc.开发了一种低成本、高灵敏度、基于荧光的检测平台， 正是这种类型的定量多抗原血清莱姆病检测。4，9，10纽约州立大学的凯迪博士使用了我们的 光栅偶联荧光等离子体（GC-FP）测定平台，以定义针对一系列 BB抗原。该初步工作表明，与STTT（90%/100%）相比，具有上级敏感性/特异性 vs 60%/100%; n=34个验证样本）和更早的结果时间。我们的方法提供了线性响应 在整个动态范围内，快速获得结果，试剂成本低。抗原点可添加用于 因此，可以以一个ELISA的价格进行更广泛的蜱传疾病（TBD）面板。 在这个第一阶段的STTR中，Ciquet将与Cady博士（纽约州立大学理工学院）和Strle博士合作 （沃兹沃斯中心）建立在我们的初步自筹资金的探索莱姆血清诊断。我们的目标是 使用大量表征良好的LD样品和适当的对照来验证我们的初步结果 来自CDC和纽约州卫生部，包括使用收集的样本估计阳性时间 纵向。与此同时，Cipher旨在通过实施一个 基于色谱的系统和自动化处理，将使GC-FP成为可行的替代方法， 如STTT和MTTT，两者都是繁琐和耗时的。最终目标是提供一个 快速、易于使用且高度准确的检测，可在传统临床实验室之外使用。基于 第一阶段的成功，第二阶段的努力将优化测试莱姆病的诊断引起的 欧洲疏螺旋体菌株，并将其扩展到其他蜱传疾病。第二阶段还将允许设计 完善和启动FDA批准程序。