Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array

使用单分子场效应晶体管阵列直接生物电子检测唾液中的 SARS-CoV-2

基本信息

  • 批准号:
    10266395
  • 负责人:
  • 金额:
    $ 81.73万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 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

项目摘要

Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array Nucleic acid tests have become the gold-standard for diagnostic testing for COVID-19, usually performed in specialized laboratories. Most are based on reverse-transcription quantitative polymerase chain reaction (qRT-PCR). The time required for specimen transport and processing results in a turnaround time that is typi- cally several days. The few rapid (<1 hour) point-of-care (POC) tests are more expensive, still require sample preparation and specialized reagents, and do not have the throughput needed for population surveillance. Di- rect testing for the virus, which also reduces requirements for multiple reagents, is a necessary step to improv- ing diagnostic testing. While four such antigen tests have been approved for detection of SARS-CoV-2 based on immunoassays to the N protein, sensitivity is limited and no quantitation of viral load is possible. We will address this gap by using DiagnostikosTM, an in-development rapid POC platform for direct, real- time, multiplexed, quantitative bioelectronic detection of biomolecules that employs an all-electronic detection device that functions at the single-molecule level. These single-molecule field-effect transistors (smFETs) are arrayed on a complementary metal-oxide-semiconductor (CMOS) integrated circuit chip. Chips will interface with an envisioned USB-stick-form-factor reader device. Robust single-domain antibodies, known as nanobod- ies and immobilized on these devices, are used for sensitive detection of viral particles and viral debris. The use of multiple nanobodies for a single protein and nanobodies for different proteins in a single assay allows for significant improvements in specificity. Nanobodies will be specific for one or more of the four major struc- tural proteins in SARS-CoV-2; the nucleocapsid (N) protein engulfing the viral RNA, the spike (S) protein, the membrane (M) protein and the envelope (E) protein. No sample preparation or specialized reagents are re- quired for detection, and the device will be designed to operate with saliva, which has very recently been shown to be a reliable medium for detecting SARS-CoV-2. Individual sensor chips can be manufactured at a cost of $35. With the addition of other nanobodies, these large dense arrays can also allow detection of many pathogens in a single test. In this Direct-To-Phase-2 SBIR program we will pursue several key innovations that are required to make such a platform possible, including isolation of nanobodies for key structure proteins of SARS-CoV-2 (Specific Aim 1), development of the smFET platform for antigen detection (Specific Aim 2), development of large CMOS arrays of these smFET devices (Specific Aim 3), and verification of detection in increasingly complex samples up to and including clinical samples (Specific Aim 4). This project is a partnership between university researchers who developed the smFET technology and a venture-based start-up venture, Quicksilver Biosci- ences, spun out to commercialize smFET technology and develop smFET/CMOS arrays for molecular diag- nostic applications.
唾液中SARS-CoV-2的直接生物电子学检测 使用单分子场效应晶体管阵列 核酸检测已成为COVID-19诊断检测的金标准, 在专门的实验室里。大多数是基于逆转录定量聚合酶链反应 (qRT-PCR)。样本运输和处理所需的时间导致周转时间通常为 cally几天。少数快速(<1小时)护理点(POC)测试更昂贵,仍然需要样本 目前,该系统缺乏制备和专用试剂,不具备种群监测所需的通量。迪- 对病毒进行正确的检测,这也减少了对多种试剂的需求,是改善 诊断测试。虽然四种这样的抗原测试已被批准用于检测基于SARS-CoV-2的 在对N蛋白的免疫测定中,灵敏度是有限的,并且不可能定量病毒载量。 我们将通过使用DiagnostikosTM来解决这一差距,这是一个正在开发的快速POC平台,用于直接、真实的 采用全电子检测的生物分子的时间、多路复用、定量生物电子检测 在单分子水平上发挥作用的装置。这些单分子场效应晶体管(smFET)是 阵列在互补金属氧化物半导体(CMOS)集成电路芯片上。芯片将与 具有设想的USB棒形状因子读取器设备。强大的单域抗体,称为纳米抗体, ies并固定在这些设备上,用于病毒颗粒和病毒碎片的灵敏检测。的 在单一测定中使用用于单一蛋白质的多个纳米抗体和用于不同蛋白质的纳米抗体允许 在特异性上有显著的提高。纳米抗体将对四种主要结构中的一种或多种具有特异性, SARS-CoV-2中的结构蛋白;吞噬病毒RNA的核衣壳(N)蛋白,刺突(S)蛋白, 膜(M)蛋白和包膜(E)蛋白。无需重新进行样品制备或使用专用试剂。 需要检测,该设备将被设计为与唾液,这是最近被 证明是检测SARS-CoV-2的可靠介质。单个传感器芯片可以在 费用35元。随着其他纳米抗体的加入,这些大的密集阵列也可以允许检测许多 病原体在一个单一的测试。 在这个直接到第二阶段SBIR计划,我们将追求几个关键的创新,需要作出 这种平台是可能的,包括分离SARS-CoV-2关键结构蛋白的纳米抗体(特异性 目标1),开发用于抗原检测的smFET平台(具体目标2),开发大型 这些smFET器件的CMOS阵列(具体目标3),以及在日益复杂的环境中的检测验证 样本,包括临床样本(具体目标4)。该项目是一个合作伙伴关系, 开发smFET技术的研究人员和一家以风险为基础的初创企业Quicksilver Biosci, 该公司致力于将smFET技术商业化,并开发用于分子诊断的smFET/CMOS阵列, nostic应用程序

项目成果

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Kenneth L Shepard其他文献

Kenneth L Shepard的其他文献

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{{ truncateString('Kenneth L Shepard', 18)}}的其他基金

A Wireless micro-ECoG Prosthesis for Speech
用于语音的无线微型 ECoG 假肢
  • 批准号:
    10513407
  • 财政年份:
    2021
  • 资助金额:
    $ 81.73万
  • 项目类别:
A Wireless micro-ECoG Prosthesis for Speech
用于语音的无线微型 ECoG 假肢
  • 批准号:
    10490475
  • 财政年份:
    2021
  • 资助金额:
    $ 81.73万
  • 项目类别:
A Wireless micro-ECoG Prosthesis for Speech
用于语音的无线微型 ECoG 假肢
  • 批准号:
    10375951
  • 财政年份:
    2021
  • 资助金额:
    $ 81.73万
  • 项目类别:
A Wireless micro-ECoG Prosthesis for Speech
用于语音的无线微型 ECoG 假肢
  • 批准号:
    10706320
  • 财政年份:
    2021
  • 资助金额:
    $ 81.73万
  • 项目类别:
Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array
使用单分子场效应晶体管阵列直接生物电子检测唾液中的 SARS-CoV-2
  • 批准号:
    10320987
  • 财政年份:
    2020
  • 资助金额:
    $ 81.73万
  • 项目类别:
Integrated, multiplexed high-frequency electronic analysis of DNA in nanopores
纳米孔中 DNA 的集成、多重高频电子分析
  • 批准号:
    8545205
  • 财政年份:
    2012
  • 资助金额:
    $ 81.73万
  • 项目类别:
Integrated, multiplexed high-frequency electronic analysis of DNA in nanopores
纳米孔中 DNA 的集成、多重高频电子分析
  • 批准号:
    8719765
  • 财政年份:
    2012
  • 资助金额:
    $ 81.73万
  • 项目类别:
Integrated, multiplexed high-frequency electronic analysis of DNA in nanopores
纳米孔中 DNA 的集成、多重高频电子分析
  • 批准号:
    8365334
  • 财政年份:
    2012
  • 资助金额:
    $ 81.73万
  • 项目类别:
Rapid Allergenic Particle Identification (RAPID)
快速过敏性颗粒识别 (RAPID)
  • 批准号:
    7337686
  • 财政年份:
    2007
  • 资助金额:
    $ 81.73万
  • 项目类别:
Rapid Allergenic Particle Identification (RAPID)
快速过敏性颗粒识别 (RAPID)
  • 批准号:
    8073325
  • 财政年份:
    2007
  • 资助金额:
    $ 81.73万
  • 项目类别:

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