A peptide-based point-of-care vertical flow assay for the rapid diagnosis of Lyme disease

基于肽的即时垂直流检测用于快速诊断莱姆病

• 批准号: 10452660
• 负责人: Paul Michael Arnaboldi
• 金额: $ 98.65万
• 依托单位: BIOPEPTIDES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-24 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452660
• 关键词: Address; Antibiotics; Antibodies; Antigen Targeting; Antigens; Bacterial Proteins; Binding; Binding Proteins; Biological Assay; Borrelia burgdorferi; Borrelia miyamotoi; Cells; Cellular Phone; Centers for Disease Control and Prevention (U.S.); Clinical; Clinical Treatment; Coupled; DBL Oncoprotein; Data; Defect; Detection; Development; Diagnosis; Diagnostic; Disease; Disease Progression; Early Diagnosis; Early treatment; Epitopes; Family; Generations; Goals; ITGB3 gene; ImProv; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Intervention; Laboratories; Laboratory Diagnosis; Legal patent; Lyme Disease; Lyme disease diagnosis; Lyme disease diagnostic; Machine Learning; Maps; Measures; Membrane; Methods; Microbe; Musculoskeletal System; Nature; Nervous system structure; OspC protein; Paper; Patient-Focused Outcomes; Patients; Peptides; Phase; Physicians; Proteins; Reader; Reporting; Research; Sampling; Serology; Signal Transduction; Small Business Innovation Research Grant; Specificity; Speed; Spottings; System; Technology; Testing; Time; Work; antibody detection; base; biological systems; blind; clinical diagnosis; cost; cost effective; cross reactivity; decorin binding protein B; design; detection assay; detection platform; diagnostic assay; efficacy testing; improved; innovation; member; multiplex detection; novel strategies; point of care; point of care testing; point-of-care detection; point-of-care diagnostics; portability; preservation; prevent; prototype; public health relevance; rapid diagnosis; rapid test; relapsing fever borrelia; screening

Abstract Project Summary There is an obvious need for new approaches and better assays for the laboratory diagnosis of Lyme disease. All attempts to develop a practical assay for the direct detection of Borrelia burgdorferi in infected patients have failed. Thus far, all proposed alternatives to serology have been unsuccessful, not cost-effective, or are still in early development. Improving serological detection in early disease is the fastest and most effective way to improve patient outcomes in Lyme disease. The antigen targets utilized in current serodiagnostic assays have considerable defects. They often contain conserved epitopes that cross-react with antibodies raised to other antigens, reducing specificity and requiring the use of a two-tier seroassay paradigm that preserves specificity at the cost of poor sensitivity in the detection of early disease. The use of peptides as serodiagnostic targets demonstrate improved efficacy, but the use of one or two peptides containing single epitopes still provides re- duced sensitivity in early disease. Putting the same old antigen targets into new platforms, no matter how in- novative the platform, will not succeed in improving serodiagnostics for Lyme disease. Both the target anti- gens and assays need an innovative approach. By incorporating unique peptides containing linear epitopes highly specific to B. burgdorferi, into a cutting-edge, multiplex, portable paper-based point-of-care diagnostic assay that uses a cost-effective smartphone-based reader, we aim to transform the diagnosis of Lyme disease. An ideal test for Lyme disease could be performed in a single step and yield an answer on the spot to support diagnosis and direct the course of clinical treatment. We have developed an innovative vertical-flow assay which allows for the multiplexed detection of IgM and IgG binding of up to 25 independent antigen targets in a point-of-care setting. The VFA design allows for uniform flow of sample across the target membrane, creating uniform binding conditions and maximizing developed signal. We coupled this design with a cost-effective portable smart-phone based reader allowing for quantitative measure of antibody binding, eliminating subjectiv- ity. By multiplexing peptide antigens each containing 1-2 epitopes unique to B. burgdorferi from multiple differ- ent antigens expressed at different stages during mammalian infection we can generate a single tier, POC as- say that can specifically and sensitively detect patient antibody at all stages of the disease. The assay can be completed in less than 25 min allowing for rapid in office results to support clinical diagnosis, thereby improving patient outcomes.

抽象的 项目摘要 莱姆病的实验室诊断显然需要新的方法和更好的检测方法。所有开发一种直接检测感染患者中伯氏疏螺旋体的实用检测方法的尝试都失败了。迄今为止，所有提出的血清学替代方案均不成功，不具有成本效益，或仍处于早期开发阶段。改善早期疾病的血清学检测是改善莱姆病患者预后的最快、最有效的方法。当前血清诊断测定中使用的抗原靶标存在相当大的缺陷。它们通常含有保守表位，可与其他抗原产生的抗体发生交叉反应，从而降低特异性，并需要使用两层血清测定范式，以保留特异性，但代价是检测早期疾病的灵敏度较差。使用肽作为血清诊断靶标可以提高疗效，但使用一种或两种含有单一表位的肽仍然会降低早期疾病的敏感性。将相同的旧抗原靶标放入新平台中，无论平台多么创新，都不会成功改善莱姆病的血清诊断。目标抗原和检测方法都需要创新的方法。通过将含有伯氏疏螺旋体高度特异性线性表位的独特肽纳入尖端、多重、便携式纸质护理点诊断检测（使用经济高效的基于智能手机的阅读器），我们的目标是改变莱姆病的诊断。莱姆病的理想测试可以一步完成，并当场给出答案，以支持诊断并指导临床治疗过程。我们开发了一种创新的垂直流检测，可在护理点环境中对多达 25 个独立抗原靶标的 IgM 和 IgG 结合进行多重检测。 VFA 设计允许样品均匀地流过目标膜，创造均匀的结合条件并最大化产生的信号。我们将此设计与基于经济高效的便携式智能手机的读取器结合起来，可以定量测量抗体结合，消除主观性。通过多重肽抗原，每个肽抗原含有 1-2 个伯氏疏螺旋体特有的表位，这些抗原来自哺乳动物感染期间不同阶段表达的多种不同抗原，我们可以生成单层 POC 检测，可以特异性、灵敏地检测疾病各个阶段的患者抗体。该检测可在 25 分钟内完成，可在办公室快速获得结果以支持临床诊断，从而改善患者的治疗结果。