Rolosense: An innovative platform for automatic mobile phone readout of active SARS-CoV-2 particles

Rolosense：用于自动手机读取活性 SARS-CoV-2 颗粒的创新平台

• 批准号: 10264612
• 负责人: Khalid S Salaita
• 金额: $ 44.97万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-21 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264612
• 关键词: 2019-nCoV; Adhesions; Affinity; Antibodies; Avidity; Binding; Biological Assay; Buffers; COVID-19 detection; COVID-19 surveillance; Caliber; Car Phone; Cellular Phone; Centers for Disease Control and Prevention (U.S.); Complementary RNA; DNA; Data; Device or Instrument Development; Diagnostic; Engineering; Environmental Impact; Flushing; Geography; Goals; Hybrids; Immobilization; Intervention; Ionic Strengths; Libraries; Ligand Binding; Mechanics; Microfluidic Microchips; Microfluidics; Motor; Mutate; Mutation; Nebulizer; Nucleic Acids; Paper; Performance; Proteins; Public Health; Publications; Publishing; RNA; Reporting; Ribonuclease H; Role; SARS-CoV-2 spike protein; Sensitivity and Specificity; Single Nucleotide Polymorphism; Specificity; Spectrophotometry; Speed; Structure; Surface; Technology; Temperature; Testing; Time; Transcend; Viral; Viral Load result; Virion; Virus; Virus Inactivation; Work; air sampling; antibody libraries; aptamer; commercialization; detection limit; experience; experimental study; high reward; high risk; improved; innovation; instrumentation; millimeter; novel; particle; prototype; response; screening; simulation; smartphone Application

Project Abstract The ultimate goal of this proposal is to develop a novel platform technology for automatic surveillance and tracing of airborne SARS-CoV-2 virus particles in real time. The centerpiece of this proposal is the “Rolosense” technology which leverages a DNA micromotor as the virus sensing and transduction material (VSTM) that can be detected by a conventional smart phone camera. This provides both geographical tracing and surveillance. Rolosense motor are comprised a DNA-coated spherical particle (5 micrometer diameter) that hybridizes to a surface modified with complementary RNA. The particle moves at speeds of over 1 micron/minute upon the addition of RNase H, which selectively hydrolyses hybridized RNA but not single-stranded RNA. DNA motors coated with virus binding ligand (VBL) stall in presence of SARS-CoV-2 virus particles. Because motors move autonomously for distances up to millimeters without intervention, the assay is fully automated, and conventional steps such as viral inactivation, RNA isolation and amplification are not required. The readout is performed using an automated smart phone app for particle tracking without the need for a spectrophotometer or fluorometer. Preliminary data shows realtime SARS-CoV-2 pseudovirus particle sensing. Milestones include the screening and identification of high affinity and high specificity VBLs. Both aptamers and antibody VBLs will be screening and validated. Simulations and experiments will be used to understand the role of temperature and environmental conditions in modulating Rolosense performance. Multivalent display of VBLs with DNA origami will enhance avidity. Finally, microfluidic chips with airborne droplet capture will be implemented and tested. The work will be performed by a highly interdisciplinary team with complementary expertise and a track-record of co-publications. PI Salaita invented the Rolosense technology and has past experience in developing cell phone diagnostics and synthetic motors. Co-I Melikian is an expert virologist, Co-I Heemstra is an expert at developing aptamers for novel targets, Co-I Ke has extensive experience in DNA origami structures for avid target binding and has co-authored work on Rolosense, Co-I Rajaraman and Primordia are experts at microfluidic device development and commercialization. Our solution offers the potential to provide an immediate solution to today’s urgent virus sensing and tracing needs.

项目摘要 这项提议的最终目标是开发一种新的自动监控平台技术 以及空气传播的SARS-CoV-2病毒颗粒的实时跟踪。这项提案的核心是 利用DNA微马达作为病毒传感和转导的“Rolosense”技术 传统智能手机摄像头可以检测到的材料(VSTM)。这既提供了 地理追踪和监视。劳斯莱斯马达由一个包裹着DNA的球形颗粒组成 (直径5微米)与互补RNA修饰的表面杂交。粒子 添加核糖核酸酶H后，移动速度超过1微米/分钟，核糖核酸酶H选择性地水解 杂化的RNA，但不是单链RNA。涂有病毒结合配体(VBL)的DNA马达 在存在SARS-CoV-2病毒颗粒的情况下。因为马达自动移动的距离可达 毫米，无需干预，检测是完全自动化的，常规步骤，如病毒 不需要灭活、RNA分离和扩增。读出是使用 自动智能手机应用程序，无需分光光度计或 荧光计。初步数据显示，SARS-CoV-2伪病毒粒子实时检测。里程碑 包括高亲和力和高特异性VBL的筛选和鉴定。适配子和适体 将对抗体VBL进行筛选和验证。模拟和实验将被用来理解 温度和环境条件在调节罗氏性能中的作用。 用DNA折纸多价展示VBL将增强亲和力。最后，微流控芯片具有 将实施和测试空中液滴捕获。这项工作将由一位高度 具有互补专业知识和联合出版记录的跨学科团队。皮萨莱塔 发明了Rolosense技术，并且过去有开发手机诊断和 合成马达。Co-I Melikian是病毒学家，Co-I Heemstra是研究 新靶标的适体，Co-i Ke在DNA折纸结构方面具有丰富的经验，可用于狂热的靶标 BINGING并与人合著了Rolosense，Co-I Rajaraman和Primordia是 微流控器件的开发和商业化。我们的解决方案提供了提供 立即解决当今紧迫的病毒检测和跟踪需求。