SBIR-STTR Award

Direct to Phase II

Crimean-Congo Hemorrhagic Fever (CCHF) is a tick-borne disease with high mortality rates in humans and highoutbreak potential. Found in 30 countries across Europe, Asia, and Africa, CCHF has an extremely widespread andgrowing range. Its causative agent, CCHF virus (CCHFV), is listed among the urgently concerning pathogensprioritized in the WHO R&D Blueprint. With at least seven regional clades, CCHFV detection by gold standard qPCR-based methods has been encumbered by significant strain-associated genetic variability. Existing tests for CCHFVare often limited to regional use, presenting a barrier to standardization and quality assurance. The WHO has calledfor the development of universal CCHFV diagnostics as a global priority. Aldatu Biosciences has pioneered the useof PANDAA technology, which enables probe-based qPCR for target detection in highly variable genomic regions bysimultaneously adapting and amplifying diverse templates. PANDAA uniquely mitigates the presence of target-proximal polymorphisms to allow otherwise divergent templates to be detected with consensus fluorescent probeswith similar sensitivities. As such, PANDAA-enabled qPCR is an ideal solution for universal detection of pathogenswith significant strain, lineage, and/or sub-type sequence diversity. Aldatu Biosciences is uniquely positioned todeliver a rapid pan-lineage qPCR-based CCHFV diagnostic. PANDAA has been successfully applied to subtype-independent detection of more than fifteen drug resistance mutation (DRM) targets in HIV. Recently, we completeddevelopment of the first pan-lineage assay for Lassa fever virus (LASV), another WHO priority pathogen with highoutbreak potential. We propose to leverage the unique capabilities of PANDAA to develop a rapid, sensitivemolecular diagnostic assay for CCHFV detection, and the first with pan-lineage coverage, through the followingspecific aims: (1) development of a pan-lineage PANDAA-CCHFV assay, leveraging proven techniques andproprietary PANDAA reagent design; (2) analytical validation of the PANDAA-CCHFV assay, including confirmationof clade inclusivity and high specificity; (3) thermostabilization of the PANDAA-CCHFV assay, in order to meet therequirements of diagnostics targeted to LMICs; (4) clinical validation of the lyophilized PANDAA-CCHFV assay, usingclinical samples representing a broad variety of circulating clades and geographies; and (5) multi-site evaluation ofthe PANDAA qDx CCHFV test kit at reference labs at CDC- and WHO-affiliated partner institutions. As the first pan-lineage assay, PANDAA-CCHFV will provide a rapid, standardized testing option for all regions within the broadrange of CCHFV. This novel, pan-lineage detection assay could ultimately be deployed in any endemic region onpre-existing qPCR equipment in central labs, and/or integrated into a closed, point-of-care system with sampleprocessing to radically improve the CCHFV diagnostic workflow.

Public Health Relevance Statement:
Crimean-Congo Hemorrhagic Fever Virus (CCHFV) is listed among the urgently concerning pathogens prioritized in the WHO R&D Blueprint. With at least seven regional clades, CCHFV detection by gold standard qPCR-based methods has been encumbered by significant strain-associated genetic variability. We propose to leverage the unique capabilities of PANDAA, a qPCR-enabling technology that mitigates the effects of clade variability on test performance, to develop a rapid, sensitive molecular diagnostic assay for CCHFV detection, and the first with pan- lineage coverage, PANDAA-CCHFV will provide a rapid, standardized testing option for all CCHF-endemic regions and can be deployed pre-existing qPCR equipment in central labs to radically improve the CCHFV diagnostic workflow.

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