Contract Award for Topic 388:Development of DNA Aptamer-Based Lateral Flow Assays for Liver Flukes (Phase I)

主题 388 的合同授予：基于 DNA 适体的肝吸虫侧向层析检测的开发（第一阶段）

• 批准号: 10022020
• 负责人: John Bruno
• 金额: $ 30万
• 依托单位: NANOHMICS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-16 至 2020-06-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10022020
• 关键词: Affinity; Antigens; Award; Biological Assay; Buffers; C-terminal; Cellular Phone; China; Clonorchis sinensis; Computer software; Contracts; Cytolysis; DNA; DNA Sequence; Data Reporting; Detection; Development; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Extracellular Domain; Fasciola hepatica; Gold Colloid; Lateral; Link; Membrane Proteins; N-terminal; Opisthorchis viverrini; Parasites; Peptides; Phase; Proline; Proteins; Quantum Dots; Reader; Recombinants; Reporter; Resources; Testing; Thailand; Visual; aptamer; base; cross reactivity; egg; magnetic beads; protein aminoacid sequence

Nanohmics proposes to develop sensitive and specific DNA aptamer-based colloidal gold sandwich lateral flow (LF) test strips from commercially available recombinant surface proteins and N-terminal peptides of Clonorchis sinensis and Opisthorchis viverrini antigens. Actual liver flukes for testing will be obtained through established contacts in China and Thailand. If the N-terminal sequence peptides do not prove to be adequate targets in Phase 1, then Nanohmics will develop aptamers in Phase 2 against C-terminal peptides and proline-rich bend regions of the sequenced trematode proteins which are likely to mark accessible extracellular domains. The same aptamers can be attached to magnetic beads and used to concentrate trematode eggs or worms from fecal slurries followed by parasite lysis or protein extraction and LF detection. And if necessary, highly fluorescent quantum dot-reporter aptamers will be employed to enhance visual detection sensitivity. In Phase 2, Nanohmics will also develop its own smart phone-controlled LF test strip reader and software applications for data reporting via the cell phone network in resource-limited environments.

Nanohmics公司建议从商业上可获得的华支睾吸虫和间日疟原虫抗原的重组表面蛋白和N端肽开发敏感和特异的基于DNA适配子的胶体金夹心横向流动(LF)测试试条。用于检测的实际肝吸虫将通过在中国和泰国建立的联系人获得。如果N端序列多肽在第一阶段被证明不是足够的靶标，那么NanoHMICS将在第二阶段针对被测序的吸虫蛋白的C末端多肽和富含脯氨酸的弯曲区开发适配子，这些区域可能标记可访问的胞外区域。同样的适配子可以附着在磁珠上，用于从粪便浆液中浓缩吸虫虫卵或蠕虫，然后进行寄生虫裂解或蛋白质提取，并进行LF检测。如果有必要，将使用高荧光量子点报告适体来提高视觉检测灵敏度。在第二阶段，NanoHmics还将开发自己的智能手机控制的LF测试条读取器和软件应用程序，用于在资源有限的环境中通过手机网络报告数据。