Rapid Diagnostic Test for Respiratory Syncytial Virus by Digital Nanobubbles
数字纳米气泡对呼吸道合胞病毒的快速诊断测试
基本信息
- 批准号:10155417
- 负责人:
- 金额:$ 50.06万
- 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:2020-05-01 至 2025-04-30
- 项目状态:未结题
- 来源:
- 关键词:AddressAffectAntibioticsAntibodiesAntiviral TherapyAreaBar CodesBindingBiological AssayBiosensing TechniquesBiosensorBypassCell Culture TechniquesCessation of lifeChildChildhoodChimeric ProteinsClinicalClinical SensitivityColorCommunicable DiseasesConsumptionCoupledCouplingDataDetectionDevelopmentDevice or Instrument DevelopmentDevicesDiagnosisDiagnosticDiagnostic ProcedureDiagnostic SensitivityDiagnostic testsDiseaseEvaluationFormulationFutureGenerationsGoldHealth care facilityHealthcareImmunoassayInfantInterdisciplinary StudyLaboratoriesLasersLateralLeadMeasuresMembrane ProteinsMethodsMicrospheresNanosphereNucleic AcidsOpticsOutcomePerformancePhysiologic pulsePneumoniaPolymerase Chain ReactionPreparationProcessProteinsQuantum DotsRapid diagnosticsReadingReagentRespiratory syncytial virusRodSamplingSecondary toSensitivity and SpecificitySignal TransductionSpecificitySpecimenSpeedSurfaceSystemTestingTimeVaccine TherapyVirionVirusWorkabsorptionanalogantibody conjugatebaseclinical translationcomputerized data processingcostdesigndetection limitdiagnostic assaydigitalgenetic strainimprovedinnovationisothermal amplificationlaboratory experiencelateral flow assaynanoGoldnanobubblenanoparticlenanorodnanoscalenoveloperationpediatric patientsperformance testsplasmonicspoint-of-care diagnosticsprototyperesearch clinical testingrespiratory pathogenresponsesensorsolid statesuccesssuperinfectionviral detectionvirus culture
项目摘要
ABSTRACT
Respiratory syncytial virus (RSV) is one of the most common causes to pediatric death globally. Rapid RSV
diagnostics is important for judicial use of antibiotics, to reduce disease spread in healthcare facilities, and to
enable prompt treatment since several RSV antiviral therapies are on the horizon. Current diagnostic methods
rely on time-consuming laboratory-based tests including virus culture and polymerase chain reaction (PCR), and
rapid diagnostic tests (e.g., lateral flow immunoassay, LFA) are not sufficiently sensitive as standalone diagnosis.
Therefore, there is an unmet need for rapid and ultrasensitive diagnostic tests for RSV. The plasmonic coupling
assay is a rapid colorimetric diagnostic test that makes use of the optical response of gold nanoparticles (AuNPs)
during the process of target recognition to analyze its concentration. Despite its easy operation, the sensitivity of
the plasmonic coupling assay is limited. In this proposed work, we aim to substantially improve the limit of
detection (LOD) of the plasmonic coupling assay by innovative digital nanobubble detection. Specifically, we
propose to directly detect intact RSV particles with antibody-conjugated AuNPs that recognize the RSV surface
fusion (F) protein. AuNPs bind to multiple RSV F proteins and lead to plasmonic coupling. Ultrashort laser pulse
selectively activates coupled AuNPs due to their enhanced absorption compared with a single AuNP. This
greater optical absorption leads to nanoscale cavitation bubbles, i.e. nanobubbles, which can be measured easily
from the bubble-induced and transient scattering. Single nanobubble generation leads to a sensitive digital
detection with “on” and “off” signals. Our preliminary results suggest ~3 orders of magnitude improvement of
LOD in detecting RSV. In this proposed work, we will firstly innovate the AuNP probe by optimizing the AuNP
formulation (size, concentration and conjugation), investigating the non-spherical nanoparticle for more efficient
virus binding, and exploring the asymmetric antibody-coated Janus nanoparticles for controlled binding. Next,
we will design and build a prototype device for automated sample loading, reading, and data processing for the
digital nanobubble assay. Lastly, we will test this assay with clinical RSV isolates (from diverse genetic strains)
and clinical specimens. By comparing with current PCR and rapid diagnostic tests, we will establish the clinical
sensitivity and specificity for the digital nanobubble test. This test is fast (< 30 minutes), low-cost (AuNP reagent
cost is similar to lateral flow immunoassay), and highly sensitive and specific for RSV. Furthermore, the direct
detection of virus particles eliminates the need of extensive sample preparation such as nucleic acid extraction.
Success of our project will meet urgent demands of rapid and sensitive RSV diagnostics and address a major
healthcare need for pediatric patients that are affected by RSV.
摘要
呼吸道合胞病毒(RSV)是全球儿科死亡的最常见原因之一。快速RSV
诊断对于抗生素的司法使用,减少疾病在医疗机构中的传播,
由于几种RSV抗病毒疗法即将问世,因此能够迅速治疗。目前的诊断方法
依赖耗时的实验室检测,包括病毒培养和聚合酶链反应(PCR),以及
快速诊断测试(例如,侧流免疫测定(LFA)作为独立诊断不够灵敏。
因此,对于RSV的快速和超灵敏诊断测试存在未满足的需求。等离子体耦合
金纳米颗粒(AuNPs)是一种快速的比色诊断测试,它利用了金纳米颗粒(AuNPs)的光学响应。
在目标识别的过程中,分析其浓度。尽管它操作简单,
等离子体偶联测定是有限的。在这项拟议的工作中,我们的目标是大大提高限制
通过创新的数字纳米气泡检测来检测等离子体偶联测定的LOD。我们特别
提出用识别RSV表面的抗体缀合的AuNP直接检测完整的RSV颗粒
融合(F)蛋白。AuNP结合多种RSV F蛋白并导致等离子体偶联。超短激光脉冲
与单一AuNP相比,由于其增强的吸收,选择性地激活偶联的AuNP。这
更大的光吸收导致纳米级空化气泡,即纳米气泡,其可以容易地测量
气泡引起的瞬态散射单个纳米气泡生成导致敏感的数字
通过“开”和“关”信号进行检测。我们的初步结果表明~3个数量级的改善
检测RSV的LOD。在这项拟议的工作中,我们将首先通过优化AuNP来创新AuNP探针,
制剂(大小,浓度和共轭),研究非球形纳米颗粒,以更有效地
病毒结合,并探索用于受控结合的不对称抗体包被的Janus纳米颗粒。接下来,
我们将设计和建造一个原型装置,用于自动样品加载,阅读和数据处理。
数字纳米气泡测定。最后,我们将使用临床RSV分离株(来自不同的遗传毒株)测试该检测试剂盒
和临床标本。通过与现有的PCR和快速诊断试验的比较,我们将建立临床上的
数字纳米气泡测试的灵敏度和特异性。该检测快速(< 30分钟),低成本(AuNP试剂
成本类似于侧流免疫测定),并且对RSV高度敏感和特异。此外,直接
病毒颗粒的检测消除了大量样品制备如核酸提取的需要。
我们的项目的成功将满足快速和敏感的RSV诊断的迫切需求,并解决一个主要的
RSV感染儿科患者的医疗保健需求。
项目成果
期刊论文数量(0)
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Zhenpeng Qin其他文献
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{{ truncateString('Zhenpeng Qin', 18)}}的其他基金
Rapid Viral Diagnostic Test by Digital Plasmonic Nanobubbles
利用数字等离子体纳米气泡进行快速病毒诊断测试
- 批准号:
10547200 - 财政年份:2022
- 资助金额:
$ 50.06万 - 项目类别:
Rapid Viral Diagnostic Test by Digital Plasmonic Nanobubbles
利用数字等离子体纳米气泡进行快速病毒诊断测试
- 批准号:
10665073 - 财政年份:2022
- 资助金额:
$ 50.06万 - 项目类别:
Rapid Diagnostic Test for Respiratory Syncytial Virus by Digital Nanobubbles
数字纳米气泡对呼吸道合胞病毒的快速诊断测试
- 批准号:
10627753 - 财政年份:2020
- 资助金额:
$ 50.06万 - 项目类别:
Rapid Diagnostic Test for Respiratory Syncytial Virus by Digital Nanobubbles
数字纳米气泡对呼吸道合胞病毒的快速诊断测试
- 批准号:
10394257 - 财政年份:2020
- 资助金额:
$ 50.06万 - 项目类别:
Optical Control of Protein Activity in Live Cells by Plasmon Assisted Light Inactivation
通过等离激元辅助光灭活对活细胞中蛋白质活性的光学控制
- 批准号:
10698186 - 财政年份:2019
- 资助金额:
$ 50.06万 - 项目类别:
Optical Control of Protein Activity in Live Cells by Plasmon Assisted Light Inactivation
通过等离激元辅助光灭活对活细胞中蛋白质活性的光学控制
- 批准号:
10223375 - 财政年份:2019
- 资助金额:
$ 50.06万 - 项目类别:
Optical Control of Protein Activity in Live Cells by Plasmon Assisted Light Inactivation
通过等离激元辅助光灭活对活细胞中蛋白质活性的光学控制
- 批准号:
10799344 - 财政年份:2019
- 资助金额:
$ 50.06万 - 项目类别:
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