Innovations in Decentralized Pan-Orthoebolavirus Diagnostics - Frequently Asked Questions (FAQs)

The 2026 Bundibugyo outbreak exposed two foundational diagnostic gaps. No validated antigen targets exist for BDBV, and no antigen RDT currently meets WHO TPP specifications for this species. Existing RDTs fail primarily at low viral loads (Ct above 30) and show inconsistent field performance even at high viral loads. Separately, clinicians currently lack tools to distinguish EVD from other febrile illnesses at first presentation, before viral load is sufficient for antigen detection.

1. Novel antigen targets and binder discovery for RDT development: Identification and characterization of BDBV-specific and pan-Orthoebolavirus antigen targets, paired with development of matched recognition elements suitable for RDT-compatible formats at clinically relevant viral loads. Eligible binder types include conventional monoclonal antibodies, synthetic nanobodies, engineered aptamers, and computationally designed scaffolds. AI/ML-based binder design approaches are strongly encouraged and should describe the computational pipeline, training datasets, and empirical validation of binding affinity, specificity, and field-relevant stability. Expected deliverables include BSL-4-validated antigen panels, matched binders, and reported limit of detection in copies per mL or TCID50 per ml. Proposals may address antigen characterization, binder development, or both. Cross-disciplinary partnerships spanning the full pipeline are preferred.
2. Host response or other biomarkers for early-illness triage: Cytokine profiles, acute-phase proteins, or other host-response signatures measurable in capillary whole blood, oral specimens, or urine that distinguish EVD from other febrile illness at or before the point of antigen detectability. Signals must be technically translatable to a field-compatible format within 3-5 years.

Applicant requirement: Access to BSL-4 containment and to outbreak patient specimens (prospective or archived, including BDBV) is required. Applicants must describe existing or formally arranged partnerships with accredited high-containment facilities and clinical sites with outbreak access. Proposals without this documentation will not be considered.

Venipuncture is slow, risky, and a bottleneck that delays diagnosis. Alternative specimen types including capillary blood, gingival-margin swabs, oral mucosal transudate, tongue dorsum swabs, and urine offer practical advantages but require rigorous, prospective analytical validation using optimal workflows before clinical adoption.

1. Venipuncture-free specimen type validation: Prospective clinical characterization of alternative specimen types for EVD diagnosis, including gingival-margin swabs, oral mucosal transudate, tongue dorsum swabs, urine, anal/rectal swab, and capillary blood, to determine at what viral load threshold each specimen type is diagnostic for EVD. Studies must use the most analytically sensitive back-end molecular assay (e.g., optimized RT-qPCR with quantitative output) to validate specimen performance and must systematically optimize and report every step in the collection-to-result workflow. For example, for swab-based specimens, this includes swab type and matrix, collection pressure, number of strokes, contact time, elution volume, stabilization and inactivation buffer, extraction approach (if any), and PCR primer-probe set. Equivalent workflow parameters should be defined and optimized for other specimen types. Results must report quantitative viral load per sample unit (e.g., target copies/swab), allowing the analytical detection limit (LOD) of the alternative specimen to be compared to viral load in paired venous blood. This goal is rigorous characterization of specimen performance to establish whether a given matrix is suitable for diagnostic use and under what conditions.
2. Inactivation workflow validation: Cross-matrix validation of chemical or physical inactivation chemistries that render BDBV, EBOV, and SUDV specimens biosafe while preserving nucleic acid and antigen integrity and enabling direct-to-PCR and RDT-based assays. Deliverables should include inactivation efficiency data from live-virus BSL-4 experiments and side-by-side assay performance data across inactivated vs. matched fresh specimens in each matrix.
3. Viral inactivation confirmation indicators: Development of a simple, integrated indicator (e.g., colorimetric, lateral flow, or electrochemical) that confirms complete viral inactivation at the point of specimen processing, without requiring BSL-4 live-virus testing for routine confirmation. Multiple mechanistic approaches are in scope.
4. Fully enclosed collection-inactivation devices: Single-use, fully enclosed devices that integrate specimen collection, inactivation, and BSL-2-safe preparation of eluate in a closed system operable by community health workers. Compatibility with capillary blood, alternative specimen types, and both downstream PCR and antigen-based assays required.

Applicant requirement: Specimen type validation (A) requires prospective access to EVD patients. Applicants must demonstrate an existing or formally arranged partnership with a clinical site in an active or recent outbreak area. Inactivation validation requires BSL-4 access for live-virus work. All other bullets may proceed at BSL-2 using validated surrogates with a BSL-4 confirmation plan. Proposals for enclosed collection-inactivation devices should be linkable to the specimen characterization data generated under Opportunity 2A and the inactivation validation data generated under Opportunity 2B. The Foundation does not expect a single applicant to span all three tracks, but proposals should describe how specimen performance data and validated inactivation chemistries will inform device design and performance specifications.

No antigen RDT currently meets WHO TPP for BDBV, the causative agent of the 2026 outbreak. Molecular confirmation requires cold-chain reagents, laboratory infrastructure, and trained operators that are unavailable in some of the most affected health zones. This Opportunity funds development of the next generation of field-ready tests.

1. Next-generation antigen RDTs with sensitivity improvements: RDTs achieving analytic sensitivity equivalent to or better than RT-PCR for Ct ≤30 for at least one Orthoebolavirus species with priority for BDBV. Sensitivity innovations may include novel antibody pairs from biomarker discovery work (Opportunity 1), signal amplification, sample concentration, or alternative detection chemistries.
2. True point-of-care molecular tests: Fully self-contained, disposable molecular platforms requiring no laboratory infrastructure, operable by health workers without specialized training, with no cold-chain requirements and result in ≤45 minutes. BDBV detection is the priority; cross-species detection is expected. Proposals using BSL-2 surrogate validation (inactivated virus, pseudovirus, recombinant RNA) at early stages are acceptable.

Applicant requirement: Proposals for RDT development should be linkable to the antigen target and binders generated under Opportunity 1. The Foundation does not expect a single applicant to span both tracks, but proposals should acknowledge how antibody discovery outputs will be sourced.

The 2026 outbreak was likely circulating in Ituri for several weeks before official declaration in May 2026. Earlier detection could have enabled earlier mobilization of support. This Opportunity funds tools that could shorten that gap in future outbreaks.

1. Sentinel surveillance tools for high-risk sites: Diagnostic or algorithmic tools enabling EVD sentinel surveillance at high-risk community sites such as health facilities, community death investigations, mining sites, border crossings, and transit hubs. Proposals may include simplified screening tools, rapid referral algorithms, or integrated alert systems.
2. Environmental and wastewater surveillance: Detection of Orthoebolavirus nucleic acid or other markers in wastewater, surface water, or other environmental matrices in endemic or high-risk areas. Proposals should address the analytical and biosafety challenges specific to high-consequence pathogen environmental surveillance, propose a sampling framework adapted to remote settings without central sewage infrastructure (e.g., mining communities, displacement camps, health facility effluent), and demonstrate or credibly project analytical sensitivity for detection prior to or concurrent with first clinical cases. Cross-sector applicants with demonstrated wastewater surveillance capability for other pathogens are encouraged to apply.
3. AI-driven early-warning systems: Data-driven tools integrating routine health system signals such as routine clinical testing results, febrile illness counts, healthcare worker absenteeism, community mortality alerts, referral patterns, and/or pharmacy utilization to generate early-warning scores for outbreak detection before laboratory confirmation. Tools must be designed for deployment on existing national health information infrastructure in hemorrhagic-fever-endemic regions, with no requirement for high-bandwidth connectivity. Cross-sector applicants with demonstrated predictive surveillance capabilities for other pathogens are encouraged to apply.

Decentralized testing is only as valuable as the quality assurance infrastructure that supports it. A persistent gap in EVD diagnostics is the absence of reference materials spanning Orthoebolavirus species, a fragmented proficiency testing landscape, and inadequate training tools for community health workers operating without laboratory supervision. The 2026 outbreak has revealed that healthcare workers often lack field-appropriate standardized instructions and training tools.

1. Reference panels and EQA materials: Development of external quality assessment (EQA) panels and commutable reference materials spanning Orthoebolavirus species (EBOV, SUDV, BDBV at minimum), clinically relevant viral load ranges, including low-positive samples at Ct ≥30, and validated across alternative specimen matrices (oral, capillary blood, urine). Materials must be usable by national reference laboratories in endemic countries without BSL-4 access, using validated inactivated or RNA-based matrices. Panels should be designed to support testing for both molecular and antigen-based assays. Panels must include high-negative controls for non-pathogenic Orthoebolavirus species or other species with cross-reactivity potential. Inclusion of these high-negative controls is required to confirm assay specificity and support appropriate clinical action when results are returned from community-level or environmental surveillance settings.
2. Field competency and efficiency tools for HCWs: Training and quality-support tools for healthcare workers and laboratory staff conducting EVD testing in decentralized or field settings, covering specimen collection, PPE procedures, inactivation, test operation, result interpretation, and result reporting. AI-powered simulation, job-aid applications, and competency assessment tools are in scope. Tools must be designed for low-literacy and low-connectivity contexts and available in French, English, and relevant local languages (Swahili, Lingala, Hema, Luganda, etc.)