PANDAA for universal, pan-lineage molecular detection of Crimean-Congo Hemorrhagic Fever virus

克里米亚-刚果出血热（CCHF）是一种蜱传疾病，人类死亡率高， 爆发潜力在欧洲，亚洲和非洲的30个国家发现，CCHF具有非常广泛的， 成长范围其病原体CCHF病毒（CCHFV）被列为急需关注的病原体之一 世卫组织研发蓝图中的优先事项。至少有7个区域分支，CCHFV检测的金标准qPCR- 的方法已经受到显著的菌株相关遗传变异性的阻碍。CCHFV的现有测试 通常仅限于区域使用，对标准化和质量保证构成障碍。世卫组织呼吁 作为全球优先事项，开发通用的CCHFV诊断方法。阿尔达图生物科学公司率先使用 PANDAA技术，该技术使基于探针的qPCR能够在高度可变的基因组区域中进行靶标检测， 同时适应和放大不同的模板。PANDAA独特地减轻了目标的存在- 允许用共有荧光探针检测其它不同模板的近端多态性 相似的敏感性。因此，PANDAA支持的qPCR是通用病原体检测的理想解决方案 具有显著的株系、谱系和/或亚型序列多样性。Aldatu Biosciences具有独特的优势， 提供快速的基于全谱系qPCR的CCHFV诊断。PANDAA已成功应用于亚型- 独立检测HIV中超过15个耐药突变（DRM）靶点。最近，我们完成了 开发了拉沙热病毒（LASV）的第一个全谱系检测方法，LASV是另一种世卫组织优先病原体， 爆发潜力我们建议利用PANDAA的独特能力，开发一种快速、灵敏的 用于CCHFV检测的分子诊断测定，并且第一个具有泛谱系覆盖，通过以下 具体目标：（1）开发一种泛系PANDAA-CCHFV检测方法，利用成熟的技术， 专有PANDAA试剂设计;（2）PANDAA-CCHFV检测的分析验证，包括确认 （3）PANDAA-CCHFV检测的热稳定性，以满足 （4）冻干PANDAA-CCHFV检测的临床验证，使用 代表各种循环分支和地理的临床样品;和（5）多中心评价 在CDC和WHO附属伙伴机构的参考实验室使用PANDAA qDx CCHFV检测试剂盒。作为第一个泛- PANDAA-CCHFV将为全球所有地区提供快速，标准化的检测选择。 CCHFV的范围。这种新的泛谱系检测方法最终可以部署在任何流行地区， 中心实验室中已有的qPCR设备，和/或集成到封闭的护理点系统中， 处理，从根本上改善CCHFV诊断工作流程。

Crimean-Congo Hemorrhagic Fever (CCHF) is a tick-borne disease with high mortality rates in humans and high outbreak potential. Found in 30 countries across Europe, Asia, and Africa, CCHF has an extremely widespread and growing range. Its causative agent, CCHF 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. Existing tests for CCHFV are often limited to regional use, presenting a barrier to standardization and quality assurance. The WHO has called for the development of universal CCHFV diagnostics as a global priority. Aldatu Biosciences has pioneered the use of PANDAA technology, which enables probe-based qPCR for target detection in highly variable genomic regions by simultaneously 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 probes with similar sensitivities. As such, PANDAA-enabled qPCR is an ideal solution for universal detection of pathogens with significant strain, lineage, and/or sub-type sequence diversity. Aldatu Biosciences is uniquely positioned to deliver 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 completed development of the first pan-lineage assay for Lassa fever virus (LASV), another WHO priority pathogen with high outbreak potential. We propose to leverage the unique capabilities of PANDAA to develop a rapid, sensitive molecular diagnostic assay for CCHFV detection, and the first with pan-lineage coverage, through the following specific aims: (1) development of a pan-lineage PANDAA-CCHFV assay, leveraging proven techniques and proprietary PANDAA reagent design; (2) analytical validation of the PANDAA-CCHFV assay, including confirmation of clade inclusivity and high specificity; (3) thermostabilization of the PANDAA-CCHFV assay, in order to meet the requirements of diagnostics targeted to LMICs; (4) clinical validation of the lyophilized PANDAA-CCHFV assay, using clinical samples representing a broad variety of circulating clades and geographies; and (5) multi-site evaluation of the 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 broad range of CCHFV. This novel, pan-lineage detection assay could ultimately be deployed in any endemic region on pre-existing qPCR equipment in central labs, and/or integrated into a closed, point-of-care system with sample processing to radically improve the CCHFV diagnostic workflow.