PHS2019-02 Omnibus Solic of the NIH, CDC, and FDA for SBIR Apps No Clinical Trial (Parent SBIR R43/4

PHS2019-02 NIH、CDC 和 FDA 的 SBIR 应用程序综合 Solic 尚未进行临床试验（母公司 SBIR R43/4

• 批准号: 10476977
• 负责人: STEVEN A BENNER
• 金额: $ 48.52万
• 依托单位: FIREBIRD BIOMOLECULAR SCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10476977
• 关键词: PHS2019; 02; Omnibus; Solic; NIH

Field Detection of arboviruses directly from mosquito traps Firebird Biomolecular Sciences, LLC. Abstract Mosquitoes are the world's deadliest creatures. In the US, mosquitoes are vectors for West Nile, many kinds of encephalitis, and viruses arriving Latin America and Africa. In the developing world, they cause millions of cases of disease and death each year. Routine insect spraying is not a "fix", as it leads to pesticide resistance, encounters public opposition, and destroys beneficial insects. Thus, the EPA, CDC, ECDPC, and their counterparts around the globe, strongly recommend surveillance before spraying. Florida law requires it. In Phase 1, we created (a) innovative technology to generate CO2 to attract mosquitoes, (b) innovative tech- nology to collect viral RNA from the attracted mosquitoes, (c) data showing that collected RNA is stable under Florida summer conditions for at least 3 days, (d) processes where captured viral RNA is amplified directly without handling mosquitos or Qiagen RNA preps, and (e) reagents that allow robust multiplex amplification. These specs, which can transform the science and practice of arbovirus surveillance, attracted TrakITNow, an innovator in instrument design. Its award-winning trap (Moskeet) counts and speciates mosquitoes, doing everything short of collecting samples from the mosquitoes and determining if these hold arboviruses. This Phase 2 project will re-configure and metric the Phase 1 innovations to allow them, in Phase 3, to be placed on Moskeet in two operational architectures (OA1 and OA2). A series of improvement and adaptations will be evaluated using tightly crafted metrics in two Aims. Meeting these metrica will trigger a commitment by TrakITNow to support Phase 3 funding to launch a combined product, Aim 1 will develop and metric Firebird's chemistry for an OA1 architecture that will convert Moskeet from a trap-count-speciate-communicate product into a trap-count-speciate-sample-communicate product. This will exploit Firebird's Phase 1 innovation (SterileQTM) that autonomously traps and stabilizes viral RNA. Different baits, RNA-stabilizing preservatives, and pre-amplification enzymes will be metricked to create, at the end of Year 1, a Moskeet-ready OA1 chemistry that allows continuous sampling of mosquitoes. Metrics include (a) >50% of infected mosquitoes leaving detectable sample, (b) >200 hours half-life of RNA, and (c) capture of >1 pg of RNA. An instrument from Insilixa (Hydra), also enhanced by Firebird's reagents, will be used for on-site virus detection. Hydra is portable, and its low complexity design allows it to be used by amateurs. Aim 2 will develop and metric Firebird's chemistry for an OA2 architecture. This will convert Moskeet into a trap-count-speciate-sample-analyze-communicate product. Here, we will develop and benchmark point-of- sampling amplification tools, specifically: (a) RT-PCR, and (b) loop amplification (LAMP). These will, as needed, incorporate reagent innovations (AEGIS, SAMRS, Biversals) to support highly multiplexed detection of arboviruses, create uniform amplification of multiple analytes, manage primer-primer products, prevent loss of PCR resources to off-target processes that create "monsters", and manage arbovirus sequence divergence.

直接从蚊虫捕捉器现场检测虫媒病毒 火鸟生物分子科学有限公司。 抽象的 蚊子是世界上最致命的生物。在美国，蚊子是西尼罗河的媒介，种类繁多 脑炎和病毒抵达拉丁美洲和非洲。在发展中国家，它们造成了数百万 每年都有病例和死亡病例。常规的昆虫喷洒并不是“解决办法”，因为它会导致农药抗药性， 遭到公众反对，消灭有益昆虫。因此，EPA、CDC、ECDPC 及其 全球同行强烈建议在喷洒前进行监测。佛罗里达州法律有此要求。 在第一阶段，我们创造了（a）产生二氧化碳来吸引蚊子的创新技术，（b）创新技术 从被吸引的蚊子中收集病毒RNA的技术，(c)数据显示收集的RNA在以下条件下是稳定的 佛罗里达夏季条件至少 3 天，(d) 直接扩增捕获的病毒 RNA 的过程 无需处理蚊子或 Qiagen RNA 制剂，以及 (e) 能够进行稳健多重扩增的试剂。 这些规格可以改变虫媒病毒监测的科学和实践，吸引了 TrakITNow（一家 仪器设计的创新者。其屡获殊荣的陷阱（Moskeet）可对蚊子进行计数和分类， 除了从蚊子身上收集样本并确定它们是否含有虫媒病毒之外，一切都完成了。 该第二阶段项目将重新配置和衡量第一阶段的创新，以便它们在第三阶段能够 放置在 Moskeet 上的两个操作架构（OA1 和 OA2）中。一系列的改进和调整 将在两个目标中使用精心设计的指标进行评估。满足这些指标将触发以下承诺： TrakITNow 支持第三阶段资金以推出组合产品， Aim 1 将为 OA1 架构开发和度量 Firebird 的化学反应，该架构将把 Moskeet 从 将陷阱计数特定样本通信产品转化为陷阱计数特定样本通信产品。这将 利用 Firebird 的第一阶段创新 (SterileQTM)，可自主捕获并稳定病毒 RNA。不同的 诱饵、RNA 稳定防腐剂和预扩增酶将在实验结束时进行计量以产生 第 1 年，Moskeet-ready OA1 化学物质，允许对蚊子进行连续采样。指标包括 (a) >50% 的受感染蚊子留下可检测样本，(b) >200 小时的 RNA 半衰期，以及 (c) 捕获 >1 RNA pg。 Insilixa (Hydra) 的仪器也经过 Firebird 试剂的增强，将用于现场 病毒检测。 Hydra 是便携式的，其低复杂性设计使其可供业余爱好者使用。 Aim 2 将为 OA2 架构开发并衡量 Firebird 的化学反应。这会将 Moskeet 转换为 陷阱-计数-特定-样本-分析-交流产品。在这里，我们将开发和基准测试点- 采样扩增工具，具体为：(a) RT-PCR 和 (b) 环路扩增 (LAMP)。这些将如 根据需要，结合试剂创新（AEGIS、SAMRS、Biversals）来支持高度多重检测 虫媒病毒，对多种分析物进行均匀扩增，管理引物-引物产品，防止损失 PCR 资源用于产生“怪物”的脱靶过程，并管理虫媒病毒序列分歧。