SBIR-STTR Award

Over 2 billion people live in areas that are affected by the transmission of arthropod-borne viruses (arboviruses) such as the flaviviruses dengue (DENV) and Zika (ZIKV), and the alphavirus chikungunya (CHIKV). These arboviruses are pathogenic and cause millions of infections annually. While the surveillance is critical to understanding infection rates, controlling outbreaks, and informing patient therapeutic efforts, there is no FDA- approved method to simultaneously detect multiple arboviral infections at the point-of-care and analyze, document, and report data in real-time. In this Phase 1 proposal we will optimize the 3Plex, a miniaturized point- of-care lateral flow test for finger prick blood or serum that can distinguish between infections of DENV, ZIKV, and CHIKV, and integrate our patented phone-based image recognition application with geo-referenced applications (e.g. District Health Information System 2, DHIS2) for real-time epidemiological reporting. Our proposed 3Plex is a low-cost test that can be done in the field without moving parts, or the necessity of trained personnel, expensive equipment, or a centralized laboratory. Studies proposed here will leverage our recent advances, including identification of monoclonal antibodies (mAbs) that recognize flaviviral non-structural protein 1 (NS1)- or alphaviral envelope (E)-specific antigen with optimal binding capacity and without cross-reactivity when utilized in a single, multiplex test. The intellectual property of the diagnostic technology being optimized in this SBIR was developed by the investigators and global rights are exclusively held by E25Bio. In Aim 1, we will fine tune a panel of immunological characterization assays focusing on mAb specificity and sensitivity and scale up production of candidate monoclonal antibodies. In Aim 2, we will advance development and validation of the 3Plex prototype. We will test DENV, ZIKV, and CHIKV supernatants from infected Vero cells, and confirm the limit of detection with de-identified human samples. In Aim 3, we will integrate our mobile phone image recognition application with predictive algorithms to report data in real-time. This project is innovative because it is the first platform that allows point-of-care arboviral antigen-based detection and real-time data analytics for infection geotagging and outbreak management. Upon successful completion of this Phase I project, we will have developed a first-of-its-kind platform, which provides an all-in-one solution to arboviral detection and data reporting, that will be ready for field testing and regulatory validation. These results will lead to a Phase II project to conduct clinical trials in Colombia to assess diagnostic efficacy in clinical settings.

Public Health Relevance Statement:
To monitor viral infections and control emerging outbreaks, a diagnostic test that can simultaneously detect among multiple viruses at the point-of-care, and a platform to report data in real-time is essential. E25Bio will generate the first lateral flow antigen-based test to simultaneously detect dengue, Zika, and chikungunya in less than 10 minutes (Aims 1 and 2), and develop a mobile smartphone application for test result data reporting (Aims 3). This viral detection and data reporting platform will be a simple yet powerful tool at the disposal of both infected individuals and organizations responsible for outbreak management.

Project Terms:
accurate diagnosis; Acute; Advanced Development; Aedes; Affect; Affinity Chromatography; Africa; Alphavirus; Americas; Antibodies; Antigens; Arbovirus Infections; Arboviruses; Area; Arthropod Vectors; base; Binding; Biological Assay; Blood; Brazil; Car Phone; Care Technology Points; Cellular Phone; Centers for Disease Control and Prevention (U.S.); chikungunya; Chikungunya virus; Clinical; Clinical Data; Collaborations; Colombia; Communication; Conduct Clinical Trials; cost; cross reactivity; Culicidae; Cultured Cells; Data; Data Analyses; Data Analytics; Data Reporting; Database Management Systems; Dengue; Dengue Infection; Detection; Diagnostic; Diagnostic tests; Disease Outbreaks; Dot Immunoblotting; effective therapy; Enzyme-Linked Immunosorbent Assay; Epidemiologic Monitoring; Epidemiology; Epitope Mapping; Equipment; Evaluation; field study; Fingers; Flavivirus; Flow Cytometry; Future; Generations; Gold; Government; health data; Health Information System; Human; Human Resources; Image; Immunologics; Individual; Infection; Infection Control; innovation; Intellectual Property; Laboratories; Lateral; Legal patent; Malaria; Methods; miniaturize; Monitor; Monoclonal Antibodies; neuropathology; new technology; Nonstructural Protein; off-patent; open source; pathogen; Pathogenicity; Patients; Performance; Phase; point of care; point-of-care diagnostics; pre-clinical; prediction algorithm; Preparation; Process; Production; prototype; Puerto Rico; Reporting; Research Personnel; Resources; Rights; Sampling; scale up; Sensitivity and Specificity; Serological; Serum; skills; Small Business Innovation Research Grant; smartphone Application; Southeastern Asia; Technology; Telephone; Test Result; Testing; Therapeutic; Time; tool; Training; transmission process; Tropical Disease; United States Food and Drug Administration; Validation; Vero Cells; Viral; viral detection; Virus; Virus Diseases; Virus-like particle; Zika Virus

Over 2 billion people live in areas that are affected by the transmission of arthropod-borne viruses (arboviruses) such as the flaviviruses dengue (DENV) and Zika (ZIKV), and the alphavirus chikungunya (CHIKV). These arboviruses are pathogenic and cause millions of infections annually. While the surveillance is critical to understanding infection rates, controlling outbreaks, and informing patient therapeutic efforts, there is no FDA- approved method to simultaneously detect multiple arboviral antigens at the point-of-care and analyze, document, and report data in real-time. In Phase I, we developed non-multiplexed tests for pan-DENV nonstructural protein 1 (NS1), ZIKV NS1, and CHIKV envelope (E) (SimPlex) and a 3-in-1, multiplexed test for all three viral antigens (3Plex). These tests were evaluated in the laboratory and using archived, de-identified patient samples. Additionally, we developed a mobile smartphone application to image capture test results, geo- locate where the image was captured, and upload the data to a central server for epidemiological mapping. In Phase II, we propose to finalize development of the SimPlex and 3Plex tests and enhance our digital platform for real-time metadata reporting to support therapeutic intervention and outbreak management. Our proposed SimPlex and 3Plex tests are low-cost, point-of-care tests that can be done in the field without moving parts, or the necessity of trained personnel, expensive equipment, or a centralized laboratory. In Aim 1, we will scale up production of lead monoclonal antibodies specific for DENV, ZIKV, and CHIKV antigens. In Aim 2, we will optimize SimPlex and 3Plex tests for whole blood detection of DENV, ZIKV, and CHIKV antigens. In Aim 3, we will produce multiple lots of SimPlex and 3Plex tests under GMP for pre-clinical evaluation using prospectively collected samples in Colombia and Brazil. In Aim 4, we will develop and informatic healthcare databased for fast integration of SimPlex and 3Plex test result data with electronic health record repositories; the digital platform will be evaluated in Colombia and Brazil. This project is innovative because it is the first platform that allows point-of-care arboviral antigen-based detection and real-time data analytics for infection geotagging and outbreak management. Upon successful completion of this Phase II project, we will have developed a first-of- its-kind platform, which provides an all-in-one solution to arboviral detection and data reporting, that will be ready for regulatory validation.

Public Health Relevance Statement:
Narrative To monitor viral infections and control emerging outbreaks, a diagnostic test that can simultaneously detect among multiple viruses at the point-of-care, and a platform to report data in real-time is essential. E25Bio Inc. will generate a first-of-its-kind lateral flow antigen-based test to simultaneously detect dengue, Zika, and chikungunya and integrate it with an evidence-based epidemiological and clinical decision support system. This viral detection and data reporting platform will be a simple, yet powerful tool at the disposal of infected individuals and their healthcare providers and organizations responsible for outbreak management.

Project Terms:
Absorbent Pads; accurate diagnosis; Acute; Aedes; Affect; Alphavirus; Antibodies; Antigens; Arboviruses; Archives; Area; Arthropod Vectors; Bar Codes; base; Bedside Testings; Biological Assay; Blood group antigen S; Brazil; Buffers; Car Phone; Care Technology Points; Characteristics; chikungunya; Chikungunya virus; Clinical; clinical decision support; Clinical Decision Support Systems; Colombia; commercialization; cost; cross reactivity; Culicidae; Data; Data Analytics; data dictionary; Data Reporting; Databases; Dengue; design; Detection; Development; Diagnostic; Diagnostic tests; digital; Disease Outbreaks; Electronic Health Record; Enzyme-Linked Immunosorbent Assay; Epidemiology; Equipment; Evaluation; evidence base; FDA approved; Feasibility Studies; field study; Flavivirus; Funding; Gold; Health; health care service organization; Health Personnel; Healthcare; Human; Human Resources; Image; Incidence; Individual; Infection; infection burden; Infection Control; infection rate; Informatics; innovation; Instruction; interoperability; Laboratories; Lateral; Lead; Metadata; Methods; Mexico; Monitor; Monoclonal Antibodies; National Institute of Allergy and Infectious Disease; new technology; Nonstructural Protein; novel; pathogen; Pathogenicity; Patients; Performance; Phase; Plasma; point of care; point-of-care diagnostics; preclinical evaluation; Preparation; Process; Production; prospective; prospective test; prototype; Recombinants; Reporting; repository; Reproducibility; Resources; Reverse Transcriptase Polymerase Chain Reaction; Sampling; scale up; Serological; Serum; Site; skills; Small Business Innovation Research Grant; smartphone Application; Source; System; Telephone; Test Result; Testing; Therapeutic; Therapeutic Intervention; Time; tool; Training; transmission process; Validation; Viral; Viral Antigens; viral detection; Virus; Virus Diseases; Whole Blood; ZIKA; Zika Virus