Isolating Viral Particles from Whole Blood

从全血中分离病毒颗粒

• 批准号: 8244067
• 负责人: MARK A HAYES
• 金额: $ 6.94万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-07 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8244067
• 关键词: Acute; Address; Behavior; Biological; Biological Assay; Biosensor; Blood; Buffers; Caring; Cells; Clinic; Clinical; Cold Chains; Collection; Complex; Cost Analysis; Data; Dengue Virus; Detection; Developing Countries; Development; Device Designs; Devices; Diagnosis; Diagnostic; Diagnostic Procedure; Elements; Goals; Health; Hemagglutination; Human Coronavirus OC43; Infection; Infectious Agent; Influenza; Kinetics; Laboratories; Liquid substance; Location; Methods; Microfluidics; Modeling; Monitor; Phase; Physiological; Procedures; Reagent; Research; Reverse Transcriptase Polymerase Chain Reaction; Saliva; Sampling; Serological; Serotyping; Structure; System; Testing; Theoretical model; Time; Translating; Venipunctures; Viral; Virion; Virus; Whole Blood; base; cost effective; design; electric field; improved; influenza virus strain; innovation; lithography; molecular recognition; particle; pathogen; respiratory; respiratory virus; viron; virus identification

DESCRIPTION (provided by applicant): Diagnostic approaches providing identification of viruses directly from bio-fluids with improved figures of merit (fast, accurate (selective, sensitive), simple, low power, and cost-effective) are needed. A new, innovative micro-fluidic strategy that can contribute to this goal is presented here. The system can rapidly and selectively separate, isolate and concentrate viruses from bio-fluids for direct identification or further assessments (immuno- or geno-recognition). The strategy is based on DC insulator gradient dielectrophoresis (DC-iGDEP) which provides not only the advantage of truly unique and non-linear separation of bioparticles, but also can remove unwanted components that are often present in complex biological samples and interfere with subsequent assays. The approach can fuse location to identification via electric field manipulation of bio-particles, thus avoiding a number of issues with current methods that require prior molecular recognition elements and commonly cold-chain reagents. The basis for the approach is a combination of dielectrophoretic and electro-kinetic forces in a single channel. The long-term objective is to integrate DC-iGDEP into a simple, cost effective, reliable biosensor that will be a component of a diagnostic platform that can be used in the clinical laboratory and ideally, amenable for surveillance and diagnosis in developing countries. As poof-of-concept, we will use dengue viruses as the target model for a blood borne virus and two respiratory viruses, human coronavirus OC43 and influenza that are typically only present in respiratory secretions and not in blood during acute infections. Dengue viruses are one of the most significant emerging infectious pathogens today and newly emerged corona viruses and influenza strains remain as important health concerns, thus the test samples will have "real world" relevance. Standard virological assays, including virus titer, hemagglutination (HA) and inhibition (HAI), RT-PCR, and other well established serological diagnostic methods will be used in parallel with the platform development to monitor sensitivity, accuracy and overall feasibility of DC-iGDEP. The goal is to isolate/concentrate/separate virus particles from typical venipuncture and respiratory sample volumes. Once developed, the approach can be modified for a broad range of medically important viruses. PUBLIC HEALTH RELEVANCE: Developing an ability to isolate and concentrate viruses from biofluids (saliva, CSF, blood) using gradient dielectrophoresis. This can be developed into devices which can detect virus at earlier phases of infection providing for better care and reduced spread of infectious agents.

描述（由申请人提供）：需要直接从具有改进功绩数字（快速，准确，敏感），简单，低功率和成本效益的生物流体鉴定病毒鉴定的诊断方法。这里提出了一种可以为此目标做出贡献的新创新的微富裕策略。该系统可以快速，有选择地分离，分离和浓缩病毒与生物流体，以直接鉴定或进一步评估（免疫或Geno-complition）。该策略基于直流绝缘体梯度介电性（DC-IGDEP），它不仅提供了真正独特和非线性分离生物颗粒的优势，而且还可以去除经常在复杂的生物样品中存在的不需要的成分并与后续的分析进行交流。该方法可以通过生物粒子的电场操作融合位置以识别识别，从而 避免了许多需要先前分子识别元件和通常冷链试剂的方法的问题。该方法的基础是单个通道中介电练习力和电动力的组合。长期目标是将DC-IGDEP整合到一个简单，具有成本效益，可靠的生物传感器中，该生物传感器将成为诊断平台的组成部分，该平台可用于临床实验室，理想情况下，可以在发展中国家进行监视和诊断。作为概念po，我们将使用登革热病毒作为血液传播病毒和两种呼吸道病毒的靶向模型，即人冠状病毒OC43和流感，通常仅在呼吸道分泌物中，而不存在于急性感染期间的血液中。登革热病毒是当今最重要的新兴病原体之一，新出现的电晕病毒和流感菌株仍然与重要的健康有关，因此测试样本将具有“现实世界”的相关性。标准病毒学测定，包括病毒滴度，血凝（HA）和抑制作用（HAI），RT-PCR以及其他良好建立的血清学诊断方法，将与平台开发同时使用，以监测DC-IGDEP的灵敏度，准确性和整体可行性。目的是分离/浓缩/单独的病毒颗粒与典型的静脉穿刺和呼吸样品体积。一旦开发，该方法就可以针对多种医学重要的病毒进行修改。 公共卫生相关性：使用梯度介电​​性产生的能力从生物流体（唾液，CSF，血液）中分离和浓缩病毒的能力。这可以发展为可以在感染的早期阶段检测病毒的设备，从而提供更好的护理和减少感染剂的传播。