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Multiplex point-of-care test for diagnosis, prognosis and serology of COVID19

用于 COVID19 诊断、预后和血清学的多重即时检测

基本信息

批准号：
10417262
负责人：
Ashutosh Chilkoti
金额：
$ 48.75万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-03 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10417262
关键词：
2019-nCoV Address Adsorption Antibodies Antigens Archives Area Biological Assay Biological Markers Blood Blood specimen Buffers CCL7 gene COVID-19 COVID-19 detection COVID-19 diagnosis COVID-19 patient CXCL10 gene Cells Clinical Collection Contact Tracing Custom Data Detection Deterioration Development Devices Diagnosis Drops Early Diagnosis Ensure Exposure to Fibrin fragment D Fluorescence Force of Gravity Geometry Glass Glycols Gold Hand Health Home Human Image Immunoassay Individual Injections Intelligence Interleukin-10 Interleukin-6 Intervention Label Length Liquid substance Methacrylates Microfluidics Molds Monitor Noise Nose Nucleocapsid Nucleocapsid Proteins Patient Care Patient Monitoring Patients Performance Pharyngeal structure Plasma Polymethyl Methacrylate Population Prevalence Printing Procedures Prognosis Proteins Quarantine Reagent Recombinants Reproducibility Resistance Risk Running SARS-CoV-2 antibody SARS-CoV-2 antigen SARS-CoV-2 infection Sampling Scanning Secure Sensitivity and Specificity Serology Serology test Serum Severe Acute Respiratory Syndrome Severities Signal Transduction Spottings Surface Swab Symptoms TNF gene Testing Text Thick Time Validation Viral Viral Antigens Whole Blood antibody detection antigen detection assay development cloud platform cost detection limit detector epidemiology study fight against fluorescence imaging high risk high throughput screening imaging platform lateral flow assay manufacturing process multiplex detection nasal swab oropharyngeal swab performance tests point of care testing point-of-care diagnostics prognostic rapid diagnosis receptor binding sample archive screening tool

项目摘要

ABSTRACT The objective of this proposal is to develop and validate a new set of low-cost, easy-to-use, point-of-care tests (POCTs) that can simultaneously detect multiple biomarkers within 30 minutes with high sensitivity and specificity to enable early diagnosis, prognosis, and serology of COVID19 patients. The multiplexed D4-POCT employs antibodies for antigen detection in the diagnosis and prognosis chips, and antigens for antibody detection in the serology chip. Capture and detection antibodies will be used to generate assays for S1 and N antigens from nasal/throat swab collection media for diagnosis, D-dimer, IL-2R, IL-6, IL-10 and TNF-α in human blood or serum for prognosis, and S1, N, and RBD viral antigens will be used as capture and detection reagents in a double- antigen serological assay to detect anti-SARS-CoV-2 antibodies in human blood and serum. Optimal reagents for assay development will be identified by leveraging the multiplexing capabilities of the D4-POCT to perform high-throughput screening and pairing of patient derived antibodies and viral antigens. The D4-POCT has three components. The first component—the D4 chip—is fabricated by inkjet printing microspots of capture and fluorescently labeled detection reagents (antibodies for antigen detection or antigens for antibody detection) directly on a protein- and cell-resistant poly(oligoethylene glycol methacrylate) brush that is grafted from the surface of glass, which reduces background noise, yielding pg/mL limit-of-detection and over 4 logs of dynamic range. The second component is a gravity-driven microfluidics cassette that encases the D4 chip and is fabricated by low-cost and high-throughput injection molding. The microfluidics cassette reduces user exposure to potentially infectious material and minimizes user interaction, as it only requires the addition of a drop of sample into the sample inlet and two drops of wash buffer into the wash buffer inlet —thereby mimicking the procedure used to carry out lateral flow assays, the gold standard of immunoassay for field use. The chip automatically performs a timed incubation and rinse step that removes the sample and any unbound detection reagent from the surface of the D4 chip, leaving the central area of the D4 chip ready for imaging by the third component—the D4Scope—a customized, low-cost, hand-held fluorescence detector that we have developed. The fluorescent spot intensities on the D4 chip are converted to analyte concentration by a built-in “App” on the D4Scope, and analyte concentrations and the raw data —the images— are both automatically uploaded to a secure cloud server. All components of the D4-POCT are easy to fabricate at high volumes as the chips are rapidly manufactured by inkjet printing picograms of reagents per test, the microfluidics cassette is fabricated by injection molding, and the D4Scope is assembled from off-the shelf parts. The impact of this proposal will be the development and clinical validation of POCTs for diagnosis, prognosis and serology—all built on the same platform— that can be globally deployed in the fight against COVID-19.

摘要