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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

基本信息

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
项目类别：
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

项目摘要

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.

摘要