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

PANDAA 用于克里米亚-刚果出血热病毒的通用、全谱系分子检测

• 批准号: 10377392
• 负责人: Iain James MacLeod
• 金额: $ 82.92万
• 依托单位: ALDATU BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-24 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377392
• 关键词: Africa; Asia; Biological Assay; Biological Sciences; Buffers; Centers for Disease Control and Prevention (U.S.); Clinical; Clinical Sensitivity; Consensus; Country; Crimean Hemorrhagic Fever; Crimean-Congo Hemorrhagic Fever Virus; Custom; Detection; Development; Diagnostic; Diagnostic Reagent Kits; Disease Outbreaks; Drug resistance; Ensure; Equipment; Europe; Evaluation; Fever; Fluorescent Probes; Freeze Drying; Genetic; Genetic Polymorphism; Genomic Segment; Genotype; Geography; Goals; Gold; HIV; Human; Institutes; Institution; Lassa virus; Methods; Molecular; Molecular Diagnostic Testing; Pathogen detection; Performance; Point-of-Care Systems; Positioning Attribute; RNA; Reaction; Reagent; Reproducibility; Sampling; Serum; Signal Transduction; Site; Specificity; Standardization; Techniques; Technology; Testing; Tick-Borne Diseases; Time; Validation; Viral Genome; Virus; Work; base; design; detection assay; detection limit; improved; innovation; low and middle-income countries; mortality; novel; pathogen; performance tests; priority pathogen; prototype; quality assurance; research and development; resistance mutation; thermostability; viral detection

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.

Crimean-Congo出血热（CCHF）是一种tick传播疾病，人类死亡率很高，高死亡率 爆发潜力。 CCHF在欧洲，亚洲和非洲的30个国家 /地区都有非常广泛的广泛性和 增长范围。它的病原体CCHF病毒（CCHFV）被列出了有关病原体 在WHO R＆D蓝图中优先考虑。至少有七个区域进化枝，通过黄金标准qPCR检测CCHFV - 基于明显的菌株相关遗传变异性的基于基于的方法。 CCHFV的现有测试 通常仅限于区域用途，这是标准化和质量保证的障碍。谁打电话给 为了开发通用CCHFV诊断作为全球优先事项。 Aldatu Biosciences率先使用了 Pandaa Technology，它可以通过基于探测的QPCR来通过高度可变的基因组区域中的靶标检测 同时适应和放大不同的模板。 Pandaa独特地减轻了目标的存在 近端多态性允许通过共识荧光探针检测到不同的模板 具有类似的敏感性。因此，支持熊猫的QPCR是用于普遍检测病原体的理想解决方案 具有明显的应变，谱系和/或亚型序列多样性。 Aldatu Biosciences独特地定位于 提供基于QPCR的快速pan-linege诊断。 Pandaa已成功地应用于亚型 独立检测HIV中超过15种耐药性突变（DRM）靶标。最近，我们完成了 开发Lassa Fever病毒（LASV）的第一个Pan-Linege测定法，另一个优先病原体 爆发潜力。我们建议利用熊猫的独特能力来发展快速，敏感的 通过以下 具体目的：（1）开发Pan-linege Pandaa-CCHFV测定法，利用良好的技术和 专有熊猫试剂设计； （2）对Pandaa-CCHFV测定法的分析验证，包括确认 进化枝的包容性和高特异性； （3）为了满足熊猫-CCHFV测定的恒温 针对LMIC的诊断要求； （4）使用冻干的熊猫-CCHFV测定法的临床验证 代表各种循环进化枝和地理的临床样本； （5）多站点评估 pandaa QDX CCHFV测试套件在CDC-和WHO相关合作伙伴机构的参考实验室。作为第一个泛 谱系测定法，Pandaa-CCHFV将为广阔的所有地区提供快速，标准化的测试选项 CCHFV范围。这个小说的泛linege检测测定法最终可以在任何地方性地区部署 中央实验室中已有的QPCR设备和/或集成到封闭的护理系统中，带有样品 处理以从根本上改善CCHFV诊断工作流程。